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BELFAST TRUST
LABORATORIES (BTL)
USER MANUALS
(Updated 5 April 2016)
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FOREWORD
Belfast Trust Laboratories are part of the Belfast Health & Social Care Trust and
incorporate all of the laboratories from Belfast City Hospital, Royal Group of
Hospitals, Mater Hospital, Musgrave Park Hospital, North & West and South East
Districts. We provide routine, emergency and regional services to parent Trusts, to
General Practitioners and to other laboratories throughout Northern Ireland and
beyond. We have strong academic links and a vigorous commitment to research and
development.
This manual has been collated from individual manuals of the constituent laboratory
departments with the aim of presenting a single overall guide to the use of the
laboratories. It is hoped that it will provide the laboratory user with a background to
the services available, list the services available within each department, provide
advice on the appropriateness of requests and help maintain the efficient use of
resources for both the laboratory and its users.
We welcome comments from users about either the manual or the services currently
available, which would lead to either improvement of the service or the manual.
MISSION STATEMENT
Belfast Trust Laboratories will provide routine diagnostic, specialist and regional
laboratory services, underpinned by teaching, research and audit to the highest
quality standards. The Laboratories will endeavour to remove all barriers to the
provision of best value services to its commissioners and clinical end users.
SERVICE AGREEMENT
Each request accepted by Belfast Trust Laboratories for examination(s) shall be
deemed to be an agreement by the user for the Belfast Health & Social Care
Laboratory services, or other accredited laboratories as may be used by Belfast
Health & Social Care Laboratories to perform testing outside repertoire, to carry out
the necessary testing and reporting function. It also implies an acceptance of the
conditions of preparation and transport as outlined in this manual.
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CONFIDENTIALITY OF SERVICE USER INFORMATION
Belfast Trust Laboratories adheres to the Belfast Health & Social Care Trust Policy
on the Data Protection Act 1998 and Protection of Personal Information (Reference
TP026/08), which outlines the legal requirement for both the Trust and its’ staff to
treat personal information confidentially and ensure all information is held securely.
COMMENTS/COMPLAINTS PROCEDURE
Belfast Trust Laboratories adheres to the Belfast Health & Social Care Trust ‘Policy
and procedure for the management of complaints and compliments’ (Copies of the
policy are available upon request from the laboratory or via the Trust Intranet).
We aim to provide high quality services. If you have a comment, compliment or
complaint about one of our services, please let us know by contacting a member of
Laboratory
staff
who
will
try
to
help
resolve
the
issue.
CONTACT DETAILS
Email: firstname.surname@belfasttrust.hscni.net
Belfast Trust Laboratories
Cancer & Specialist Services, 1st Floor, Old Generator House, Lisburn Road, Belfast
City Hospital, BT9 7AB
Laboratory Director
Clinical Director Laboratories
Tissue
Pathology
&
Molecular
Services Manager
Blood Sciences Manager
Ms Anne Moffatt
Dr Clodagh Loughrey
Mr Laurence Tucker
028 95 049976
028 90 663556
028 95 049524
Mrs Patricia McKinney
078 2514 6497
Administration Services Manager
Mr Eddie Adair
028 95 049592
Quality Co-ordinator
Information Manager
Mrs Rachelle Moore
Ms Tessa Hughes
028 95 047653
028 95 047553
Nb. Contact details for each laboratory are available within the relevant
section.
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CONTENTS
VERSION
PAGES
CLINICAL BIOCHEMISTRY
6.0
5 – 68
HAEMATOLOGY
12.0
69 – 129
HISTOCOMPATIBILITY & IMMUNOGENETICS
7.0
130 – 138
IMMUNOLOGY
8.0
139 – 175
MICROBIOLOGY
6.0
176 – 221
TISSUE PATHOLOGY
9.0
222 – 256
GENETICS
6.0
257 – 284
CLINICAL 2.0
285 – 291
PNEUMATIC
SPECIMENS
TUBE
TRANSPORT
OF
TRANSPORT OF SPECIMENS TO THE LABORATORY
2.0
292 – 301
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d
Looking for specific information?
Type the “test name” or a “ key word” in the text search box on the tool bar above
(or use ‘Crtl+F’ keyboard shortcut if search box not displayed)
Department of Clinical Biochemistry
Belfast Trust Laboratories
The Department of Clinical Biochemistry provides CPA (UK) accredited services for
primary, secondary and tertiary healthcare providers in the hospitals and community
practices of most of greater Belfast and specialised services in Endocrinology,
Metabolic Investigations, Newborn Blood Spot Screening, Toxicology, Trace Metals
and Regulatory peptides for the Province.
BTL Biochemistry is also active in clinical research and maintains close working
relationships with Departments in The Queens University Belfast and University of
Ulster to facilitate innovative research and development. BTL has the primary
Northern Ireland role in the vocational training of all the professions within laboratory
medicine – Chemical Pathologists, Clinical Scientists and Biomedical Scientists –
including teaching of medical and science undergraduates.
Services are centred in the Kelvin Laboratory Building on the Royal Group Hospitals
site with additional limited facilities at the Belfast City Hospital and Mater Infirmorium
Hospitals.
This publication is intended to provide a convenient source of information to enable
users to access our services, order appropriate tests, ensure that samples are
presented to the laboratory correctly and aid interpretation and follow up of results.
Every effort is made to ensure the information provided is up to date and accurate,
however interpretation of any test result should always be in clinical context using
the reference and other information provided in the laboratory report. Any clinical or
interpretative queries should be directed to the Duty Biochemist on week days
(028906) 33798 or the on call Doctor (via the hospital switch board) out of hours.
Contact information is also given for key staff and their primary responsibilities and
they may be contacted directly as appropriate.
It is a requirement of our National Accreditation body, CPA (UK) Ltd. that all
laboratory documents are controlled this means that any hard copies made of this
information may not be valid beyond the day of printing. Web publications (Belfast
Trust Intra / Internet & GP website) will be the current version.
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Locations:
The Clinical Biochemistry Department is centred on the Royal Group Hospitals site
of the Belfast Trust in the Kelvin Laboratories building. Limited repertoire facilities are
also situated at the Mater Hospital (Laboratory building) and the Belfast City Hospital
(Tower Block laboratories on ‘C’ floor).
Telephone:
General queries should be directed to Duty Biochemist Line at the RVH (Tel: 02890
633798). A menu selection will direct you to the appropriate area for the help
required. You can also access laboratory sites or key individuals as appropriate
using the details below
RVH Kelvin Laboratories:
Central Laboratory
02890 633798 (direct)
BCH Laboratories:
‘C’ Floor Biochemistry
02895040916 (direct)
MIH Laboratory:
02895041329
(Laboratory Reception Desk)
02895041333
(Biochemistry Laboratory)
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Professional Contacts
Clinical Lead
Service
Manager
Medical
Consultants
Operational
Managers
Clinical
Scientists
Margaret McDonnell
Consultant Clinical
Biochemist
Brian Cairns
margaret.mcdonnell@belfasttrust.hscni.net
028906 33230
brian.cairns@belfasttrust.hscni.net
Dr C Loughrey
Consultant Chemical
Pathologist
Dr K Ryan
Consultant Chemical
Pathologist
Dr G Connolly
Consultant Chemical
Pathologist
Prof. Ian Young Consultant
Chemical Pathologist
clodagh.loughrey@belfasttrust.hscni.net
028950 49057
077952 71803
028950 48823
kathryn.ryan@belfasttrust.hscni.net
028950 47455
Grainne.connolly@belfasttrust.hscni.net
028906 34126
i.young@qub.ac.uk
028906 32743
Dr Brona Roberts
Consultant Chemical
Pathologist
Mr Alastair Magill
(Regional Specialist
Services)
Mr Steve Coward
(Automation)
Mrs M McDonnell
Consultant Clinical
Scientist - Endocrinology
Dr Gareth McKeeman
Consultant Clinical
Scientist - General
Biochemistry
Immunoproteins &
Oncology
Dr Jennifer Cundick
Principal Scientist
Metabolic Laboratory
Dr Kirsty Spence Principal
Clinical Scientist,
Endocrinology
Dr Lee Armstrong
Principal Clinical Scientist,
Regulatory Peptides
Dr Jenny Hamilton
Principal Clinical Scientist,
Toxicology
Brona.Roberts@belfasttrust.hscni.net
028906 35301
alastair.magill@belfasttrust.hscni.net
02890 634895
steve.coward@belfasttrust.hscni.net
02890 634446
margaret.mcdonnell@belfasttrust.hscni.net
02890 633230
gareth.mckeeman@belfasttrust.hscni.net
02890 634714
Jennifer.Cundick@belfasttrust.hscni.net
02890 633064
kirsty.spence@belfasttrust.hscni.net
02890 634043
lee.armstrong@belfasttrust.hscni.net
02890 632737
jenny.hamilton@belfasttrust.hscni.net
02890 633164
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Normal working hours:
Normal working hours are 09:00 – 17:00 hours Monday to Friday with limited routine
services on Saturdays, Sundays and Bank Holidays to 13:00 hours. At all other
times a 24 hour emergency service is available from the RVH and BCH sites. At the
weekend and Bank Holidays routine specimens should reach the laboratory before
11:00 hours.
Clinical Biochemistry out of hour’s services for the Mater Hospital are provided from
the Belfast City Hospital site (From 1st November 2013) on the following basis:
Weeknights
Saturdays/Sundays
Bank Holidays
16:00 to 08.30
12:00 to 08:30
12:00 to 08:30
Urgent Samples:
The laboratory must be telephoned to arrange all urgent samples before the
specimen is sent to the laboratory and instructions will be given. It is NOT sufficient
to mark the request form “urgent”. The requesting clinician is responsible for
arranging transport of urgent samples to the laboratory.
Transport of Specimens:
Transport of specimens to the laboratory is usually via the pneumatic tube system
where available, the hospital porters, or delivery vans as appropriate. Blood gas,
CSF, and samples on ice must NOT be transported via pneumatic tube system since
this may compromise sample integrity for these tests. Please check the relevant
hospital protocol for detailed transport arrangements at your site.
Please note the laboratory is not responsible for the maintenance or operation of the
pneumatic tube systems and does not carry any supply of PODS. Received PODS
are returned directly to correct station using the inbuilt chip.
Mater Transport Arrangements for Out of Hours (OOH) Services
Send all samples to the Mater laboratories as normal at all times, laboratory staff will
arrange transport of the samples to the Belfast City Hospital. Samples will be
transported on a 20 minute cycle. Motorcycle couriers are on duty to transport urgent
samples from Mater Laboratories to the Belfast City Hospital on weekdays during
rush hour (16:00-19:00).
If the sample is a clinical emergency and must be transported immediately, phone
the laboratory with the patient details to make arrangements. This must be conveyed
to Biomedical Scientist OOH on ext 41333 or Bleep 1706.
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Completion of Biochemistry request forms:
Except in the limited areas making use of electronic ordering all other requests for
patient testing must be using a fully completed Clinical biochemistry Test request
Form.
The importance of supplying the correct information in a legible form cannot
be over-stressed since specimens cannot be accepted for analysis where the
identifying information on either the specimen or request form is inconsistent
or inadequate.
Essential
Desirable
Sample Tube
Patients full name*
Date of birth and/or
Hospital number (or H&C
number)
Date and time
Destination for report
Request Form
Patients full name*
Date of birth and/or hospital
number (or H&C number)
Patient’s location and
destination for report
Clinical information
Date and time of sample
collection (which is
sometimes essential)
Patient’s address
Patient’s sex
Practitioner’s bleep
number or contact number
Patient’s consultant or GP
Name of requesting practitioner
* or proper coded identifier
On occasion some of the desirable data may be essential; it is preferable to supply
all of the information detailed above to ensure that the patient is not inconvenienced
or put at unnecessary risk due to delay in provision of results.
All specimens from known or potential carriers of Category III pathogens, e.g.
Hepatitis B, Hepatitis C or HIV MUST be clearly marked with hazard labels on the
request form and the specimen tube.
Laboratory results:
Hospital patient results can be accessed electronically as soon as they become
available in the Laboratory. Printed reports are also delivered to wards throughout
the day. GP reports are transmitted by electronic download (EDI) in to individual
practice systems.
The laboratory will endeavour to contact Clinical Staff about unexpected and /
or abnormal results which are identified as potentially relevant to the
immediate wellbeing of the patient concerned. A list of critical values for
telephone reporting is available on request.
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Rejection of Requests for analysis or restrictions on reporting of
results:
The Laboratory may be obliged to reject a request for analysis or restrict
reporting of results when:

There is insufficient information supplied to enable unequivocal identification
of the patient, the specimen, the tests required or the source of the request.
 The specimen has been collected inappropriately e.g. unsuitable
anticoagulant or preservative.
 The integrity of the specimen is in question e.g. leaked in transit, undue delay
in transport, a pattern of results that suggests significant error (eg. sample
from a drip arm or contaminated with EDTA).
 There is interference present which invalidates one or more test results
(excessive haemolysis, icterus or turbidity).

The source of the request will in all such eventualities be informed of the problem,
assuming that this is identifiable. Please contact the Duty Biochemist for a full
explanation of the reason for rejection or restriction of reporting and advice on how to
avoid this eventuality.
Measurement Uncertainty:
Measurement Uncertainty recognises that all results are subject to measurement
error and in order to compare a test result with a previous result or with a specific
decision value it can be useful to have a feel for the reliability of the result.
Measurement Uncertainty (MU) uses the principles of metrology to provide a
measure of the dispersion of values within which the true result is likely to lie. Please
contact the Laboratory for more details.
Turnaround Times:
Results for some emergency requests are available within minutes of the sample
arriving in the laboratory. Whenever possible, routine requests are reported within
one working day of receipt. However, the frequency and turnaround times of some
investigations are, of necessity, longer. A guide to expected turnaround times for
individual test requests is included in the following table of test guidance information.
If required, the Laboratory will endeavour to respond to a specific request more
rapidly please contact the Duty Biochemist.
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Referral Tests:
Specialised tests which are not available in the Belfast Trust may be sent to selected
referral laboratories for analysis by arrangement. The specific referral laboratory is
identified on the laboratory report. Further details are available upon request.
“Adding–on” Requests to Existing Samples:
It may be possible to retrospectively request additional testing on a sample already
in the possession of the laboratory when this would facilitate efficient clinical
management. However, this depends on the integrity of the sample in our
possession, its suitability for the additional test, its whereabouts and within the
laboratory system and current workload levels being experienced. Results may take
up to 12h to report so if urgent results are required it is always preferable to take a
fresh sample. Add-on testing remains at the discretion of the laboratory because of
the additional risks and work it involves and we would appeal to you not to abuse this
aspect of the service particularly.
Point of Care Testing:
Whilst the use of POCT devices can aid successful outcomes for patients, there is
also a significant potential for causing patient harm. The POCT Committee oversees
POCT within the Trust and the Clinical Biochemistry Department will assist with
ensuring that the analytical performance of devices and user training is adequate
and appropriately quality controlled. All applications for new or replacement devices
must be sent to the Trust POCT Committee for approval. Please contact the
laboratory for advice relating to POCT.
The POCT office can be contacted via by email DL-POCT@belfasttrust.hscni.net or
by telephone 028 90633007.
Consumables:
The following stocks will be stored and issued by the Biochemistry Laboratory.
Plasma Catecholamines
Calcitonin
Trace metals
Urinary Catecholamines, calcium, phosphate
Urinary Cortisol, Creatinine, Trace metals, Protein,
5HIAA, 5HT
Hypopak
Acylcarnitines
Pyruvate
CSF Neurotransmitters
Glass tube containing preservative
Lithium heparin tubes
Sodium heparin tubes
24 hr urine bottle with preservative
24 hr plain urine bottle
Contains all tubes required for the investigation of
hypoglycaemia
Blood spot card
Glass tube with preservative
Specific request form and tubes
The following pages provide specific information on Clinical Biochemistry
tests available including sample requirements and interpretative information.
P a g e | 12
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
ACYLCARNITINES (TMS)
GUTHRIE CARD
2-3 blood spots
Fill designated card areas with blood
Qualitative report.
Critical results are
acted on immediately
10-14 days
ACYLCARNITINE PROFILE
(TMS)
PLASMA
BLOOD
Lithium Heparin
2ml Green top
Sample sent to Referral Laboratory
Qualitative report
5-14 days
ACYLCARNITINES
(post mortem)
Contact Paediatric
Laboratory
(Tel. 02890 632148)
Sample sent to Referral Laboratory
Qualitative Report
5-14 days
ADRENOCORTICOTROPHIC
HORMONE (ACTH)
BLOOD ON ICE
4ml Purple Top EDTA
tube
Transport to lab immediately on ice
<46 ng/L at approx.
9am
7 days
ALANINE
AMINOTRANSFERASE
BLOOD
GEL
4ml Gold top SST
Male: 4–41 U/L
Female: 4-33 U/L
1 day
ALBUMIN
BLOOD
GEL
4ml Gold top SST
35 – 50 g/L
1 day
ALBUMIN CREATININE RATIO
(ACR, see MICROALBUMIN)
URINE (RANDOM)
Yellow top Monovette
or
Sterile Universal
<3.0 mg/mmoL
1day
ALCOHOL ( ETHANOL)
BLOOD
GEL
4ml Gold top SST
Not normally
detected. Reported in
mg/dL
1 day (90 min
urgent
request).
Part of a Bone and a Lipid profile.
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TEST
SAMPLE
REQUIREMENTS
ALDOSTERONE
BLOOD.
Plain and EDTA
Must send BOTH 4ml
red top and 4 ml
purple top
BLOOD
GEL
4ml Gold top SST
ALKALINE PHOSPHATASE
(ALP)
COMMENT
REFERENCE
RANGE
Expected
TAT
See lab report
1 Month
Part of Bone and Liver profiles.
Plasma concentrations of Sulfasalazine
& Sulfapyridine may lead to false
results. Contact the lab for further
information.
Adult: 30 - 130 U/L
(Age dependent.
Up to 4 times adult
upper limit during
growth)
1 day
Contact Laboratory
Qualitative report
Contact
Laboratory
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
ALKALINE PHOSPHATASE
ISOENZYMES
BLOOD
GEL
4ml Gold top SST
ALPHA-1-ACID
GLYCOPROTEIN
BLOOD
GEL
4ml Gold top SST
0.47 - 1.25 g/L
ALPHA-1-ANTITRYPSIN
BLOOD
GEL
4ml Gold top SST
Age related.
Adult range:
0.89 – 1.89g/L
P a g e | 14
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
ALPHA-1-ANTITRYPSIN (GI
protein loss)
FAECES
Plain Universal
10g (minimum of 2g
from babies)
Send to lab immediately for freezing to
prevent protein degradation
Sample then forwarded to Referral
Laboratory
ALPHA-1-ANTITRYPSIN
PHENOTYPES
BLOOD
GEL
4ml Gold top SST
ALPHA-AMINOADIPIC
SEMIALDEHYDE (A-AASA)
URINE
Minimum 3ml
Plain universal
Sample sent to Referral Laboratory
Please note- this analysis is NOT CPAaccredited.
Detailed in report
ALPHA-FETOPROTEIN (AFP)
BLOOD
GEL
4ml Gold top SST
See Appendix 5 for ranges
Contact Laboratory
for interpretation
1 day
ALPHA SUB UNIT
BLOOD
GEL
4ml Gold top SST
Sample sent to referral laboratory. Contact
laboratory for further details.
<1 IU/L.
Female mid cycle/
menopause <3 IU/L
3 Months
ALUMINIUM
BLOOD
Blue/Black top trace
metal bottle
6 mL
Special tube – contact laboratory for
supply.
<10 ug/L
2 weeks
AMIKACIN
BLOOD
GEL
4ml Gold top SST
Contact Microbiology for clinical advice.
See Belfast Trust Guidelines
for empirical antibiotic
prescribing in hospitalised
adults.
Trough Level <5mg/L
1 day
4 weeks
Qualitative report
14 days
P a g e | 15
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
AMINO ACIDS
BLOOD
EDTA
4ml Purple top
OR
Lithium Heparin
4ml Green top
Send to lab immediately.
It is important to give full clinical details
including diet and any drug treatment.
Age related
5-14 days
AMINO ACIDS
URINE
Plain Universal
Random urine (2ml)
Send to lab immediately.
It is important to give full clinical details
including diet and any drug treatment.
Qualitative report
5-14 days
AMMONIA
BLOOD
EDTA ON ICE
4ml Purple top filled
Send to lab immediately on ice.
Patient should be resting and fasting if
possible.
Prem. Neonate <200
Term neonate <100
1 day (90 min
requested
urgently).
Plasma concentrations of Sulfasalazine
& Sulfapyridine may lead to false
results. Contact the lab for further
information.
Adult:
Female
11.2-48.2 umol/L
Male
14.7 – 55.3 umol/L
AMPHETAMINE (see Drugs of
Abuse Screening)
AMYLASE
BLOOD
GEL
4ml Gold top SST
28 – 100 U/L
1 day or 90
min in
emergency
AMYLASE
URINE
Random
16 - 49 U/24hr
1 day or 90
min in
emergency
ANDROSTENEDIONE
BLOOD
Plain
4ml Red top bottle
Pre-pubertal (0.5 –
9yrs) <0.8nmol/L
Post -pubertal
2.0–5.4nmol/L
21 days
Gel tube unsuitable.
P a g e | 16
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
ANGIOTENSIN CONVERTING
ENZYME (ACE)
BLOOD
GEL
4ml Gold top SST
Contact Laboratory
12 – 68 U/L
4 days
Anti thyroid peroxidise
antibody
BLOOD
GEL
4ml Gold top tube
Adult <34 IU/ml
for age and
pregnancy related
reference ranges
please contact lab
1 day
APOLIPOPROTEINS
BLOOD
GEL
4ml Gold top SST
Contact Laboratory
ARGININE VASOPRESSIN
BLOOD
Plastic Heparin on
ice.
Must be received in the lab within 30
minutes Sample sent to a Referral
Laboratory.
ARSENIC
BLOOD
Blue/black topped
trace metal bottle
6mL
Patient must not have taken any form of
seafood or red wine for 5 days to exclude
non-toxic forms of arsenic.
Lp(a)
<30mg/100ml
Apo A1
Male : 104 –
202mg/dL
Female: 108225mg/dL
ApoB
Male: 66-133mg/dL
Female: 60 –
117mg/dL
3 Months
< 130 nmol/L
2 weeks
P a g e | 17
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
ARSENIC
URINE
Random urine.
Plain, plastic
universal
25 mL.
Patient must not have taken any form of
seafood or red wine for 5 days to exclude
non-toxic forms of arsenic.
Unexposed
< 40 nmol / mmol
Creatinine
2 weeks
Metal bedpan must NOT be used to collect
urine.
Occupational
exposure
< 173 nmol / mmol
Creatinine
BLOOD
LITHIUM HEPARIN
ON ICE
4ml Green top tube
Send to Laboratory on ice within 30
mins of taking sample
Normal >32 umol/L
ASCORBATE
14 day
Risk of
Vit C deficiency
10 – 32 umol/L
Severe risk of Vit C
deficiency < 10
umol/L
ASIALOTRANSFERRIN
(Tau Transferrin)
Nasal fluid
Minimum 0.2 mls
Plain glass or plastic
Sample sent to referral laboratory.
For analysis of ? CSF leak
ASPARTATE
AMINOTRANSFERASE (AST)
BLOOD
GEL
4ml Gold top SST
Markedly sensitive to haemolysis. See
liver profile.
Plasma concentrations of Sulfasalazine
& Sulfapyridine may lead to false
results. Contact the lab for further
information.
BARBITURATE SCREEN
(See Drug Screen – Serum
5 - 40 U/L
1 day
P a g e | 18
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
BARIUM
URINE
Random urine.
Plain, plastic
universal
25 mL.
Metal bedpan must NOT be used to collect
urine.
< 1.5 ug/L
2 weeks
Not normally detected
1 Week
Male: 0.1-1.6 umol/L
Female: 0 –1.1umol/L
14 days
0.8 – 2.2mg/L
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
0 – 300 ug/L
4 weeks
BATTENS DISEASE ENZYMES
BENCE-JONES PROTEIN
Contact Laboratory Sample sent to
Referral Laboratory
URINE
Qualitative analysis:
EMU in a plain
universal
Quantitative analysis:
24hr urine bottle, no
preservative.
See Drug Screen – Serum
BENZODIAZEPINE SCREEN
BETA CAROTENE
BETA-2-MICROGLOBULIN
BETA-2-MICROGLOBULIN
BLOOD
GEL
4ml Gold top tube
BLOOD
GEL
4ml Gold top tube
Analysed with vitamins A and E
URINE
Plain universal
Send to lab immediately for stabilising
Sample then sent to Referral Laboratory
P a g e | 19
TEST
SAMPLE
REQUIREMENTS
BILE SALTS PROFILE
(FOR THE DIAGNOSIS OF
BILE ACID BIOSYNTHESIS
DISORDERS)
BILIRUBIN, TOTAL
BILIRUBIN, Direct
BIOTINIDASE
BLOOD GASES
REFERENCE
RANGE
Expected
TAT
0 – 14 U/L
2days
See Carbohydrate – deficient
transferrin
BETA TRACE PROTEIN,
BETA-2 TRANSFERRIN
BILE ACIDS
COMMENT
GEL
4ml Gold top tube
BLOOD
Lithium Heparin
2ml Green top
OR
2 mls urine in a plain
tube
BLOOD
GEL
4ml Gold top SST
BLOOD
GEL
4ml Gold top SST
BLOOD
Lithium Heparin
2ml Green top
BLOOD
Heparinised syringe
Sample sent to Referral Laboratory
Sample sent to Referral Laboratory.
N.B. This test is not for assessment of
obstetric cholestasis - see ‘BILE ACIDS’
4-6 weeks
<21 umol/L
1 day
< 5 umol/L
1 day
Sample sent to Referral Laboratory
2.5 – 10.5 U/L
10-14 days
Ensure there are no air bubbles in the
sample. Remove needle and cap syringe.
Send to lab immediately. Sample must be
analysed within 45 minutes.
Do not send via pneumatic chute.
Do not send in ice.
pH:7.35 – 7.45
30 mins
pC02: 4.5 – 6.0 kPa
p02: 11.0 – 14.0kPa
Tot. C02: 22-29
mmol/L
Std. Bicarb: 22-26
mmol/L
BE: -2.5 - +2.5
P a g e | 20
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
BONE PROFILE
BLOOD
GEL
4ml Gold top SST
See individual
analytes
1 day or 90
min in
emergency
BONE MARKERS
(RESORPTION: βCROSSLAPS, FORMATION:
BONE SPECIFIC ALP)
BRAIN NATRIURETIC
PEPTIDE (NTProBNP)
BLOOD
GEL
4ml Gold top SST
Fasting sample. Must reach laboratory
within 3 hours.
Age/Sex related
reference intervals.
BLOOD
GEL
4ml Gold top SST
<125 ng/L excludes HF as a cause of
signs/symptoms
Age-matched
reference intervals
2 days
C3
BLOOD
GEL
4ml Gold top SST
0.75 - 1.65 g/L
2 days (MonFri) If
received after
2pm on Fri,
will be
analysed on
Mon
C4
BLOOD
GEL
4ml Gold top SST
0.14 - 0.54 g/L
2 days (MonFri) If
received after
2pm on
Friday, will be
analysed on
Monday
P a g e | 21
TEST
SAMPLE
REQUIREMENTS
CA 125
BLOOD
GEL
4ml Gold top SST
CA 19-9
BLOOD
GEL
4ml Gold top SST
CADMIUM
CADMIUM
REFERENCE
RANGE
Expected
TAT
0 – 35 U/mL
24 hrs
Contact Laboratory
0 – 37 U/mL
24 hrs
BLOOD
Blue/black topped
Trace Elements bottle
6mL
Special tube – contact laboratory for
supply.
Non-smokers
< 27 nmol/L
Smokers
< 54 nmol/L
2 weeks
URINE
Random urine.
Plain, plastic
universal
25 mL.
BLOOD
Plain Heparinised ON
ICE (no gel)
4ml
Metal bedpan must NOT be used to collect
urine. Contact laboratory for bottles.
< 10 nmol / mmol
Creatinine
2 weeks
Must be sent to lab on ice immediately.
Males < 8.4 ng/L
Females <5 ng/L
2-4 weeks
CALCIUM
BLOOD
GEL
4ml Gold top SST
Avoid stasis when taking sample.
2.10 – 2.60 mmol/L
1 day
CALCIUM
URINE
24hr urine collection
in bottle with acid
preservative
Contact laboratory for special bottle
containing acid.
2.5 – 7.5 mmol/24hr
CALCITONIN
COMMENT
P a g e | 22
TEST
SAMPLE
REQUIREMENTS
COMMENT
CALCULI
Plain Universal
Send to lab in sterile container
REFERENCE
RANGE
Expected
TAT
1 week
CANNABANOIDS
See Drugs of Abuse
Screening
CARBAMAZEPINE
BLOOD
Trough sample
Plain tube without gel.
4ml
Single drug regime:
4-12 mg/L Multiple
drug regime: 4-8mg/L
CARBOXYHAEMOGLOBIN
BLOOD
EDTA Heparinised
(no gel)
4ml
Tube must be filled to exclude air. Patients
treated with oxygen prior to sampling may
show normal levels even after severe
poisoning.
Non-smokers: 0.51.5%
Average smokers: 45%
Heavy smoker: 8-9%
CARCINOEMBRYONIC
ANTIGEN (CEA)
BLOOD
GEL
4ml Gold top SST
Contact Laboratory
Non-smokers: 0-4
ng/ml
Smokers: 0-10 ng/ml
CAROTENE
see Vitamins A&E
28 hr
14 days
CARBOHYDRATE-DEFICIENT
TRANSFERRIN
CATECHOLAMINES
28 hr
SEE ASIALOTRANSFERRIN (CSF leak)
and TRANSFERRIN GLYCOFORM
ANALYSIS (Congenital Disorders of
Glycosylation).
NB- Testing for assessment of alcohol
intake NOT available.
BLOOD (plasma)
By special arrangement only. Contact
laboratory
14 days
P a g e | 23
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
CATECHOLAMINES
(Adrenaline, Noradrenaline &
Dopamine)
URINE
24hr urine bottle with
acid preservative
Contact laboratory for details
Adrenaline
5 – 120 nmol/24hr
Please see Appendix 6 for
investigations of Phaeochromocytoma
and Paraganglioma.
Expected
TAT
Noradrenaline
50 – 560 nmol/24hr
Dopamine
300 – 3900nmol/24hr
CATECHOLAMINES
PAEDIATRIC
URINE
Contact laboratory for details
Universal containing
acid
Random urine sample
containing acid
Age related reference
range printed with
results.
7 days
CERULOPLASMIN
BLOOD
GEL
4ml Gold top SST
Adult:
0.16 - 0.45 g/L
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
CHLORIDE
BLOOD
GEL
4ml
95 – 108 mmol/L
1 day
Samples received after 2 pm on a Friday
will be separated, frozen and analysed on
Monday. If sending a sample from outside
BHSCT please transfer promptly to allow
analysis within 24 hours.
P a g e | 24
TEST
SAMPLE
REQUIREMENTS
CHLORIDE OUTPUT
COMMENT
REFERENCE
RANGE
Expected
TAT
URINE
24h collection
110-250 mmol/24hr
1 day
CHOLESTEROL
BLOOD
GEL
4ml Gold top SST
<5.0 mmol/L
1 day
CHOLINESTERASE
(Plasma, Phenotyping)
BLOOD
EDTA
4ml Purple Top Tube
Contact laboratory Sample sent to Referral
Laboratory
CHROMIUM
Blood.
Blue/black topped
trace metal bottle.
6mL.
Special tube – contact laboratory for
supply.
< 30 nmol/L
MHRA action limit
134 nmol/L
2 weeks
CHROMIUM
URINE
Random urine.
Plain, plastic
universal
25 mL
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE.
< 2.0 nmol/mmol
creatinine
2 weeks
CHROMOGRANIN A
BLOOD
EDTA or Lithium
Heparin bottle
4ml Purple top or
green top
Transported on ice to Biochemistry
Laboratory
0 – 3 nmol/L
7-10 days
CHOLECALCIFEROL
(See Vit. D)
1 month
P a g e | 25
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
CICLOSPORIN
BLOOD
EDTA
4ml Purple top
Trough sample
100 – 250 ug/L (renal
transplant patients)
Same day if
sent before
12 noon
CITRATE
URINE
24hr urine bottle with
acid preservative
24hr urine collection
1.6 – 4.5 mmol/24hrs.
1 month
COBALT in
Whole Blood
Blood.
Blue/black topped
trace metal bottle.
6mL.
Special tube – contact laboratory for
supply.
5 - 30 nmol/L
MHRA action limit
119 nmol/L
2 weeks
COBALT in
Plasma
Blood.
Blue/black topped
trace metal bottle.
6mL.
Special tube – contact laboratory for
supply.
0.8 – 8.0 nmol/L
2 weeks
COBALT
URINE
Random urine.
Plain, plastic
universal
25 mL.
METAL BED PAN MUST NOT BE USED
TO COLLECT URINE.
< 3.0 nmol/mmol
creatinine
2 weeks
COCAINE (see drugs of abuse
screening)
P a g e | 26
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
COPPER
BLOOD
Plain or plastic
heparinised tube
(orange cap) or
blue/black topped
Trace Elements bottle
6 mL
Special tube – Contact Laboratory for
supply.
Adults
12.6 – 26.7 umol/L
1 week
Children
0-3 mnths 1.5-7
umol/L
4-6 mnths 4-17
umol/L
7-12 mnths 8-20.5
umol/L
1-13 yrs 12.5-22
umol/L
Pregnancy
27-50 umol/L
< 0.60 umol/24hr
2 weeks
COPPER
URINE
24 hr urine bottle
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE.
COPPER
Liver biopsy
Place the tissue inside a plain, plastic
Universal bottle. Do NOT put the biopsy
onto tissue paper or metal foil.
< 50 ug/g dry weight
4 weeks
CORTISOL
BLOOD
GEL
4ml Gold top SST
Diurnal variation. Note time of sample on
request form.
171-536 nmol/L 7-10
am
1day
64-327 nmol/L
pm
CORTISOL
URINE
24hr urine bottle
24hour urine collection. MUST be in a
plain bottle ie no preservative
< 210 nmol/24hr
4-8
14 days
P a g e | 27
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
C-PEPTIDE
BLOOD
Gel
4ml Gold top SST
Must be received in lab within 4 hrs
(Serum must be separated from cells AND
FROZEN within 4 hours)
1.1-4.4 ug/L (fasting)
7 days
C-REACTIVE PROTEIN
BLOOD
GEL
4ml Gold top SST
<5 mg/L
1 day
CREATINE KINASE
BLOOD
GEL
4ml Gold top SST
Male: 40 – 320 U/L
Female: 25 – 200 U/L
1 day or 90
min in
emergency
CREATINE KINASE
ISOENZYME
ELECTROPHORESIS
BLOOD
GEL
4ml Gold top SST
CREATININE
BLOOD
GEL
4ml Gold top SST
For Information on AKI ALERT System
and GAIN Acute Kidney Injury Protocol
see Appendix 1.
40-110 umol/L
URINE
24hr plain urine bottle
Therapeutic concentrations of the
following drugs can cause artificially
low creatinine results: Adrenaline,
Noradrenaline, Dopamine, Rifampicin,
Levodopa and Calcium dobesilate.
7-18 mmol/24hr
BLOOD & URINE
GEL
24hr plain urine bottle
4ml Gold top tube
Need to take account of body surface area
when dealing with paediatric samples.
71-151 mL/min
CREATININE CLEARANCE
4 weeks
1 day
1 day
P a g e | 28
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
CRYOGLOBULINS
3 x 6 ml blood
samples into three
pre-heated (37 oC)
plain clotting sample
tubes; 1 x 4ml blood
sample into a preheated (37oC) EDTA
sample tube.
A further “room
temperature” plain
clotted sample should
also be taken.
These tubes are
supplied by the
laboratory.
Requesting cryoglobulin analysis: For
outpatient appointment please contact
Angela Hurley (02890 630003) or Angela
McCusker (02890 630004) at the
Immunology Day Centre. Patients will be
booked into level 6B Outpatients for blood
collection. Appointment will also be
confirmed with laboratory staff who supply
the necessary pre-heated blood collection
equipment to level 6B.
Detected or not
detected.
7 days
CSF Analysis
(Oligoclonal bands)
Paired CSF (~2mL)
and one Gold top gel
sample tube must be
submitted together
CSF SPECTROPHOTOMETRY
(XANTHOCHROMIA)
CSF No. 3 or 4
sample in Plain
universal; min
volume 500 uL &
BLOOD
GEL
4ml Gold top SST
If detected the
cryoglobulin is
quantitated and
classified.
If there is a genuine reason that the patient
cannot attend this clinic it may be
necessary to arrange collection using taxi
transportation, please contact laboratory
02890 632624.
All Royal Hospitals requests will be
collected from wards / clinics, please
contact laboratory 02890 632624.
Please contact laboratory BEFORE
collecting sample.
Oligoclonal bands:
not detected
14 days
Contact laboratory
1 day
P a g e | 29
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
URINE
Random sample
No preservative
Analysis of Cystine is a screening test
Qualitative
48hrs
7-DEHYDROCHOLESTEROL
BLOOD
Lithium Heparin
2ml Green top
For diagnosis of Smith Lemli Opitz
Syndrome
Sample sent to Referral Laboratory
<2 umol/L
4-6 weeks
DEHYDROEPIANDROSTERONE SULPHATE
BLOOD
Gel
4ml Gold top SST
Varies with age
Contact lab for age
related reference
ranges
1 day
DIGOXIN
BLOOD
Gel
4ml Gold top tube
Contact laboratory
0.5 – 2.0 ug/L
Target range in heart
failure 0.5 – 1.0 ug/L
1 day
CYCLOSPORIN (see
CICLOSPORIN)
CYSTINE
DIHYDROXY-TESTOSTERONE
DRUGS OF ABUSE
SCREENING Amphetamines,
Benzodiazepines,
Cannabinoids, Cocaine,
Methadone and Opiates.
Mephedrone, Tramadol,
Buprenorphine , Alcohol also
available on request.
DRUG SCREEN - SERUM
(Barbiturates, Benzodiazepines,
Tricyclics)
Sample should be taken 6-8 hrs after
dose. Hypokalaemia potentiates toxicity.
Sample sent to Referral Laboratory
URINE
Urine in a
preservative free
bottle.
30ml
BLOOD
Plain
4 mL Red topped
tube
3 Months
Screening
results: 24 hr.
If
confirmation
required
5days.
Gel tube unsuitable
1 day
P a g e | 30
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
ELASTASE, PANCREATIC
FAECES
Plain universal
Random
Excessively liquid samples may be
unsuitable
Normal 200 - >500
ug/g stool
4 weeks
Mod. Pancreatic
insufficiency: 100 –
200 ug/g stool.
Severe pancreatic
insufficiency: <100
ug/g stool.
ELECTROLYTE PROFILE
BLOOD
GEL
4ml Gold top SST
See individual
analytes
Estimated Glomerular
filtration Rate.
Calculated in
electrolyte profile
>60mL/min
ETHYLENE GLYCOL
BLOOD
Plain
4ml
FERRITIN (see Iron Profile)
BLOOD
GEL
4ml Gold top tube
By special arrangement only. Contact
Laboratory
1 day or 90
min in
emergency
Not normally detected
1 day
Male: 30 – 400 ug/L
1 day
Female, pre
menopausal: 13 150ug/L
Female: post
menopausal: 13 –
300 ug/L
P a g e | 31
TEST
SAMPLE
REQUIREMENTS
FOLATE
FOLLICLE STIMULATING
HORMONE (FSH)
COMMENT
REFERENCE
RANGE
Expected
TAT
BLOOD
GEL
4ml
4.6 – 18.7 ug/L
1 day
BLOOD
GEL
4ml Gold top tube
Male:
1.5 – 12.4 U/L
2 days
Female:
Follicular 3.5-12.5 U/L
Ovulatory 4.7-21.5
U/L
Luteal 1.7-7.7 U/L
Postmenopausal
25.8 -134.8 IU/L
FREE ANDROGEN INDEX
CALCULATION
FEMALES:
20-49yrs 0.34 –
3.87%
≥50yrs
2.38%
FREE LIGHT CHAINS
BLOOD
GEL
4ml Gold top SST
2 days
0.16 –
Kappa 3.3-19.4 mg/L 24 hrs MonLambda 5.7-26.3mg/L Fri. If
K:λ ratio 0.26-1.65
received after
2pm on
Friday, will be
analysed on
Monday
P a g e | 32
TEST
SAMPLE
REQUIREMENTS
FREE T4
REFERENCE
RANGE
Expected
TAT
BLOOD
GEL
4ml Gold top SST
12-22 pmol/L
for age and
pregnancy related
reference ranges
please contact lab
1 day
FREE T3
BLOOD
GEL
4ml Gold top SST
1 day
GADOLINIUM
PLASMA
Blue/black topped
Trace Metal Bottle
6ml
URINE
Plain universal
2ml
3.1-6.8 pmol/L
for age and
pregnancy related
reference ranges
please contact lab
<0.5ug/L
GALACTITOL
GALACTOKINASE
Contact Metabolic
Laboratory
(tel 02890 632148)
GALACTOSE-1-PHOSPHATE
BLOOD
Lithium heparin green
top 2ml
COMMENT
Specimen should be taken >96 hrs after
administration of gadolinium containing
contrast media
Sample sent to Referral Laboratory.
Qualitative Report
Levels are raised in Classical
Galactosaemia and Galactokinase
deficiency.
Can be used to exclude these, even if
patient has had a blood transfusion. Also
used for monitoring treatment.
Sample sent to Referral Laboratory
N.B. Urine galactitol used as first line
screen
Sample sent to Referral Laboratory.
Urgent transit required.
Send Mon-Thurs AM only (to arrive in
laboratory before 1pm).
Contact laboratory (tel 02890 632148)
Treated
Galactosaemic
(non fasting):
<0.6umol/g Hb
2 weeks
P a g e | 33
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
GALACTOSE-1-PHOSPHATE
URIDYL TRANSFERASE (RED
BLOOD CELL)
BLOOD
EDTA (no gel)
0.5ml
Blood transfusion invalidates result.
Contact laboratory before sending (tel
02890 632148).
This test is qualitative screen for Classical
Galactosaemia.
Quantitative sample can be sent to referral
laboratory by prior arrangement if required.
Qualitative report
1-5 days
GAMMA GLUTAMYL
TRANSPEPTIDASE
BLOOD
GEL
4ml Gold top SST
Male: 10-71 U/L
Female: 6-42U/L
1 day
GASTRIN
BLOOD
EDTA ON ICE
4ml Purple top
BLOOD
GEL
4ml Gold top SST
EDTA blood (fasting if possible)
transported on ice to laboratory
0 – 40 pmol/L,
0-100 ng/L
7-10 days
Contact Microbiology for clinical advice.
See Belfast Trust Guidelines
for empirical antibiotic
prescribing in hospitalised
adults.
EDTA blood transported on ice to
laboratory
Trough Level <1mg/L
1 day
N-terminal (GL-N)
0-70pmol/L, 0250ng/L
gall
C-terminal (GL-C)
0-45pmol/L, 0150ng/L
Random 4.0 – 8.0
mmol/L
Fasting 4.0 - 6.0
mmol/L
2-4 weeks
GENTAMICIN
GLUCAGON
BLOOD
EDTA ON ICE
4ml Purple top
GLUCOSE
BLOOD
Fluoride/EDTA
4ml Grey top tube
1 day or 90
min in
emergency
P a g e | 34
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
GLUCOSE
CSF
Plain sterile container
0.5ml
URINE
Random urine in plain
sterile container
10ml
Plasma glucose should be measured at
the same time.
75% of plasma level
1 day
If screening result is abnormal, a follow-up
blood enzyme analysis is sent to a
Referral Laboratory for diagnosis.
Qualitative report
5-14 days
BLOOD
Plain or plastic
heparinised tube
(orange cap) or
blue/black topped
Trace Elements bottle
6mL
BLOOD
GEL
4ml Gold top SST
BLOOD
Gel
4ml Gold top SST
Special tube – contact laboratory for
supply.
Therapeutic range
2.5 – 10.1 umol/L
2 weeks
NOT for hospital use
See separate
analytes
1 day
Random levels are not useful.
Contact
Laboratory
7 days
BLOOD
EDTA ON ICE
12ml Purple top tubes
Fasting sample required.
Transport to lab on ice within 2 hours or
nearest lab for separation for transport
frozen.
GLYCOSAMINOGLYCANS
GOLD
GP Profile
GROWTH HORMONE
GUT AND ISLET CELL
HORMONES
2-4 weeks
P a g e | 35
TEST
SAMPLE
REQUIREMENTS
HAPTOGLOBIN
BLOOD
GEL
4ml Gold top SST
HEAVY METAL SCREEN
comprising arsenic (see arsenic
comment), cadmium, lead,
mercury and thallium.
Blood
HEAVY METAL SCREEN
comprising arsenic (see arsenic
comment), cadmium, chromium,
cobalt, lead, mercury, nickel and
thallium.
Urine
Random urine.
Plain, plastic
universal
25 mL.
HIGH DENSITY LIPOPROTEIN
CHOLESTEROL (HDL)
BLOOD
GEL
4ml Gold top SST
Calculated value
HDL/CHOLESTEROL RATIO
Blue/black topped
trace metal bottle.
6mL.
COMMENT
REFERENCE
RANGE
Expected
TAT
0.25 – 1.77 g/L
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
Special tube – contact laboratory for
supply.
See individual
analytes for reference
ranges
2 weeks
Metal bedpan must NOT be used to collect
urine.
See individual
analytes for reference
ranges
2 weeks
>1.0 mmol/L
1 day
<5.0
HOMOCYSTEINE
BLOOD
EDTA ON ICE
4ml Purple top tube
Sample must arrive in the lab ON ICE
within 30 min.
6 – 13 umol/L
1 week
HOMOGENTISIC ACID
URINE
Plain universal
10ml
Random sample of urine, send to lab
immediately
Not normally detected
5-14 days
P a g e | 36
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
HUMAN CHORIONIC
GONADOTROPHIN (HCG)
BLOOD
GEL
4ml Gold top SST
<5.0 U/L – pregnancy
unlikely for all
women.
1 day
3-HYDROXYBUTYRATE
BLOOD
Fluoride/EDTA
1ml Grey top tube
Use sugar tube. Contact laboratory for
details (tel 02890 632148).
Please give clinical details on request,
including glucose level if available.
Interpretation
1-5 days
dependent on
concurrent plasma
glucose/clinical
setting. Contact lab
for interpretation (tel
02890 633064)
5-HYDROXY-INDOLEACETIC
ACID (5HIAA)
URINE
24hr urine collection
in plain bottle
Patient should avoid pineapple, banana,
plums, tomatoes, kiwi fruit and walnuts for
3 days before and during the collection
period
10 – 47 umol/24hr
14 days
5-HYDROXYTRYPTAMINE
URINE
24hr urine collection
in plain bottle
Can be analysed on same sample as 5HIAA. Patient should avoid pineapple,
banana, plums, tomatoes, kiwi fruit and
walnuts for 3 days before and during the
collection period.
0.30 – 1.30 umol/24hr
14 days
17-HYDROXY
PROGESTERONE
BLOOD
Plain
4ml red topped bottle,
no gel
Gel tube unsuitable.
Neonates must be >48hrs old before
taking sample.
Please contact lab immediately on the
suspicion of CAH
Males
1.8 – 10.4 nmol/L
Females:
follicular phase
0.3 -3.3 nmol/L
Luteal phase
2.9 – 15.2 nmol/L
7 days
P a g e | 37
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
Pregnancy
First Trimester
7.6- 29.6 nmol/L
Second Trimester
10.3- 25.8 nmol/L
Third Trimester
13.7-57.2 nmol/L
Please contact lab
regarding paediatric
reference range
Age related normal
ranges are given on
report
IgG subclasses (IgG 1,2,3)
4ml BLOOD
GEL
4ml Gold top SST
IMMUNOGLOBULINS
4ml BLOOD
GEL
4ml Gold top SST
INHIBIN B
Contact Laboratory
Sample sent to Referral Laboratory
INSULIN
BLOOD
Gel
4ml Gold top SST
INSULIN-LIKE GROWTH
FACTOR 1 (IGF1)
BLOOD
Gel
4ml Gold top SST
Must be received in lab within 4 hrs Serum must be separated from cells AND
FROZEN within 4 hours
Blood glucose estimation must be carried
out at the same time.
Must be sent to lab immediately.
Age related.
Adult ranges:
IgG: 6.0 – 16.0 g/L
IgA: 0.8 – 4.0 g/L
IgM: 0.5 – 2.0 g/L
7 days
7 days
4 months
2.6-24.9 mu/L
7 days
Varies with age
7 days
P a g e | 38
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
INTESTINAL
DISACCHARIDASES
JEJUNAL BIOPSY
SAMPLE
Contact Laboratory for details (tel 02890
632148).
Minimum biopsy weight of 1.5 mg
required.
IRON PROFILE
BLOOD
GEL
4ml
Contact Laboratory for details
See individual
analytes
1 day
IRON
BLOOD
GEL
4ml Gold top tube
Male: 10.6 – 28.3
umol/L
Female: 6.6 – 26.0
umol/L
1 day or 90
min in
emergency
IRON
URINE
24 hr urine bottle or
Desferrioxamine
chelation test 6 hour
collection
Metal bedpan must NOT be used to collect
urine.
Contact trace metal laboratory (90631859)
for details of the desferrioxamine chelation
test procedure
2.0 – 5.0 umol/24hr
2 weeks
IRON
Liver biopsy
Place the tissue inside a plain, plastic
Universal bottle. Do NOT put the biopsy
onto tissue paper or metal foil.
SEE INTESTINAL DISACCHARIDASES
< 1500 ug/g dry
weight
4 weeks
KEPPRA (Levetiracetam)
BLOOD
Plain
4ml Red top tube
Gel tube unsuitable.
Can be analysed on same sample as
lamotrigine if patient is on both drugs
2-46 mg/L
1 week
LACTATE
BLOOD
Fluoride/EDTA
4ml Grey top tube
Send to laboratory immediately.
0.6 – 2.4 mmol/L
1 day or 90
min in
emergency
JEJUNUM BIOPSY
P a g e | 39
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
LACTATE
CSF
Fluoride/oxalate
Use sugar tube. Send to laboratory
immediately
Adult: 1.1–2.4 mmol/L
LACTATE DEHYDROGENASE
(LDH)
(IFCC Assay)
BLOOD
GEL
4ml Gold top tube
135 - 225 U/L
1 day or 90
min in
emergency
LAMOTRIGINE
BLOOD
Plain
4ml Red top tube
Level increases markedly with haemolysis
or delayed separation.
Plasma concentrations of Sulfasalazine
& Sulfapyridine may lead to false
results. Contact the lab for further
information.
Gel tube unsuitable. Contact laboratory for
sampling times.
1 – 15 mg/L
1 week
LAXATIVES
BLOOD
Contact laboratory
LEAD
BLOOD
Any bottle with
anticoagulant
LEAD
URINE
24 hr urine bottle or
Random urine
Plain, plastic
universal
25 mL.
LIPASE
BLOOD
GEL
4ml Gold top tube
Sample sent to Referral Laboratory
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE.
Expected
TAT
1 month
Adults and children
older than 5 years
< 0.50 umol/L
Children (0-5 years)
< 0.24 umol/L
2 weeks
< 30 nmol/24hr
or
< 7 nmol/mmol
creatinine
2 weeks
13 – 60 U/L
P a g e | 40
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
LIPID PROFILE
BLOOD
GEL
4ml Gold top tube
See individual
analytes
1 day
LITHIUM
BLOOD
GEL
4ml Gold top tube
Patient should be fasting for 14hrs before
sample is taken (can be done on same
sample as biochemistry profile). For
information on optimal targets in high risk
individuals please refer to JBS 2
guidelines.
Sample should be taken 12hrs after dose.
Note: If serum Li >4.0mmol/L,
haemodialysis is required. Haemodialysis
should be considered at Li >3mmol/L if
patient is toxic as serum levels do not
reflect severity of overdose.
Prophylaxis: 0.4 – 1.0
mmol/L
Acute mania: <1.2
mmol/L
1 day
LIVER PROFILE
BLOOD
GEL
4ml Gold top tube
See individual
analytes
1 day or 90
min in
emergency
LOW DENISTY LIPOPROTEIN
(LDL)
CALCULATED in
Lipid profile
<3.0 mmol/L
1 day
LUTEINISING HORMONE (LH)
BLOOD
GEL
4ml Gold top tube
Contact Laboratory for details.
LYSOSOMAL ENZYME
SCREEN (previously known
as ‘white cell enzymes’)
BLOOD
EDTA
5ml Purple top tube
Contact laboratory for more details of tests
included in screen (tel 02890 632148). For
urgent delivery. Send Mon-Thurs AM only
(to arrive in lab before 1pm).
NB- Does not include MPS disorders
Male: 1.7 – 8.6 U/L
2 days
Female:
Follicular 2.4–12.6U/L
Ovulatory 14-95.6 U/L
Luteal
1.0-11.4
U/L
Postmenopausal
7.7 – 58.5 IU/L
4-6 weeks
P a g e | 41
TEST
SAMPLE
REQUIREMENTS
MAGNESIUM (serum)
BLOOD
GEL
4ml Gold top tube
MAGNESIUM
URINE
24hr collection in a
PLASTIC bottle
MANGANESE
MERCURY
MERCURY
METANEPHRINES
COMMENT
REFERENCE
RANGE
Expected
TAT
0.70 – 1.00 mmol/L
1 day or 90
min in
emergency
24hr collection in a PLASTIC bottle.
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE
2.4 – 6.5 mmol/24hr
1 day
BLOOD
Blue/black topped
Trace Elements bottle
6mL
BLOOD
Blue/black topped
trace metal bottle.
6mL.
Special tube – contact laboratory for
supply.
76 – 396 nmol/L
2 weeks
Patient must not have taken any form of
seafood for 5 days
< 20 nmol/L
2 weeks
URINE
24 hr urine bottle or
Random urine.
Plain, plastic
universal 25 mL.
Patient must not have taken any form of
seafood for 5 days
< 50 nmol/24 hours
or
< 5 nmol / mmol
Creatinine
2 weeks
URINE
24hr urine bottle with
acid preservative
Please see Appendix 6 for
investigations of Phaeochromocytoma
and Paraganglioma.
Contact laboratory for details
Special tube – contact laboratory for
supply.
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE
Metanephrine
141 - 1289 nmol/24h
Normetanephrine
440 - 2960nmol/24h
Methoxytyramine
198 -1680 nmol/24h
P a g e | 42
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
BLOOD
Heparinised (no gel)
4ml
Fill the tube
<1.5%
METHANOL
BLOOD
Plain
4ml Red top tube
By special arrangement only. Contact
Laboratory
Not normally detected
METHOTREXATE
BLOOD
Plain
4ml Red top tube
Note: It is ESSENTIAL to contact the lab
before therapy is started. Samples should
be taken at 24hr intervals after high dose
of therapy until serum level is <0.1umol/L.
MICROALBUMIN/CREATININE
RATIO (ACR)
URINE
Plain universal
Random urine
METHADONE (See Drugs of
Abuse Screening)
METHAEMOGLOBIN
MICRONUTRIENT SCREEN
Trace Metals
(Se, Cu, Zn)
BLOOD
Orange Li Hep tube
or blue/black topped
Trace Elements bottle
GEL
Vitamin B12, Folate, Iron
GEL, protect from
light
Vitamins A & E
Vitamin C
Lithium Heparin on
ice, protect from light
1 day
1 day
<3 mg/mmol
creatinine
Please send separate form with each
sample tube.
Expected
TAT
See individual
analytes for reference
ranges.
1 day
P a g e | 43
TEST
SAMPLE
REQUIREMENTS
MOLAR HCG
MONOPROLACTIN
COMMENT
REFERENCE
RANGE
Expected
TAT
Male: 63-245 mU/L
Female: 75-381 mU/L
3 days
Sample sent to Referral Laboratory
BLOOD
GEL
4ml Gold top tube
Monoprolactin is reported only if total
prolactin >700 mU/L
MUCOPOLYSACCHARIDES
See GLYCOSAMINOGLYCANS
MYCOPHENOLIC ACID
Sample sent to Referral Laboratory
1 week
Fasting if possible, contact Laboratory for
advice.
3-4 weeks
NEUROENDOCRINE TUMOUR
SCREEN
NEUROKININ A
BLOOD
EDTA ON ICE
12ml
BLOOD
EDTA ON ICE
NEURON-SPECIFIC ENOLASE
NEUROTRANSMITTERS (CSF)
Transported to lab on ice within 2 hours or
nearest lab for separation for transport
frozen.
Transported on ice to laboratory
Sample sent to Referral Laboratory
CSF
Contact Laboratory
for specific request
form/tubes required
and transit
requirements
(tel 02890 632148)
Sample sent to Referral Laboratory
0-20 pmol/L
0-20 ng/L
7-10 days
P a g e | 44
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
NICKEL
URINE
Random urine.
Plain, plastic
universal 25 mL
METAL BEDPAN MUST NOT BE USED
TO COLLECT URINE.
< 13 nmol / mmol
Creatinine
2 weeks
NON-ESTERIFIED FATTY
ACIDS (FREE FATTY ACIDS)
BLOOD
Fluoride/EDTA
2ml Grey top tube
Sample sent to referral lab.
Interpretation
dependent on
concurrent plasma
glucose/clinical
setting. Contact lab
for interpretation (tel
02890 633064)
2 weeks
OCCULT BLOOD
FAECES
Contact laboratory
Sample from 3
successive motions
should be taken
Patient should not eat red meat, dark fish,
or uncooked vegetables containing
peroxidises for 3 days before and during
sampling. Patient should not take iron or
Vitamin C containing medication for 2 days
prior to or during sampling
Not normally detected
24 hrs
OESTRADIOL
BLOOD
GEL
4ml Gold top tube
Male: 28-156 pmol/L
Female:
Follicular 46-607
pmol/L
Ovulatory 315-1828
pmol/L
Luteal 161-774
pmol/L
Postmenopausal:
<201 pmol/L.
Pregnancy 1st
trimester: 789-15781
pmol/L
2 days
P a g e | 45
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
OLIGOSACCHARIDES
URINE (FRESH)
Plain universal
10ml urine
Sample is sent to Referral Laboratory
Qualitative Report
3-4 weeks
URINE
Plain universal
5ml random urine
sample
Give full clinical details including diet and
drug treatment. Do not collect in Boric acid
container as this makes analysis
impossible.
See ALPHA-1-ACID GLYCOPROTEIN
Qualitative report
5-14 days
URINE
Plain universal
10ml random urine
sample
NB. Boric Acid
container NOT
suitable
BLOOD
GEL
4ml Gold top
Age related
5-14 days
275 – 295 mmol/kg
1 day or 90
min in
emergency
URINE
Plain universal
20ml random urine
sample
URINE
24hr urine in
specialised bottle
containing acid.
Random urine for
children
250 – 1250 mmol/kg
1 day or 90
min in
emergency
Male:
0.08 – 0.4mmol/24hr
1 week
OPIATES( See Drugs of Abuse
Screening)
ORGANIC ACIDS (ORG)
OROSOMUCOID
OROTIC ACID
OSMOLALITY
OSMOLALITY
OXALATE
Contact Laboratory for details.
Female: 0.04 – 0.32
mmol/24hr
P a g e | 46
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Children:
<1 year: 0.015 – 0.26
mmol/mmol creat
1-5 years: 0.011 –
0.12 mmol/mmol
creat
5-12 years: 0.0059 –
0.15 mmol/mmol
creat.
>12 years: 0.0021 –
0.083 mmol/mmol
creat.
0-50 pmol/L,
0-200 ng/L
Expected
TAT
PANCREATIC POLYPEPTIDE
BLOOD
EDTA ON ICE
4ml
Take fasting if possible, transported on ice
to laboratory
PANCREATIC HORMONE
SCREEN
BLOOD
EDTA ON ICE
12ml
BLOOD
GEL
4ml
Take fasting if possible, transported on ice
to laboratory
PARAQUAT
Blood
Lithium Heparin
10ml
Sample taken >4 hours after overdose
Not normally detected
1 day
PARAQUAT
URINE
Plain universal
30ml
Sample taken >4 hours after overdose
Not normally detected
1 day
PARACETAMOL
7-10 days
2-4 weeks
NB. New treatment line joining >100mg/L
Therapeutic levels at
at 4h to >50mg/L at 12h; valid for a single 4hr: 5-12 mg/L
OD at a known time. Staggered ingestions
or uncertain timing information indicates
treatment regardless of level.
P a g e | 47
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
PARATHYROID HORMONE
(PTH)
BLOOD
EDTA
4ml
15 - 65 pg/ml
4 days
PARATHORMONE-RELATED
PEPTIDE (PTHrP)
Contact Laboratory.
Special tubes
required
NO ICE REQUIRED. MUST BE
RECEIVED IN LAB WITHIN 12HRS OF
VENAPUNTURE
Request serum calcium at same time on a
separate sample.
Sample sent to referral Laboratory by prior
arrangement. Contact
90633230/90634043
PHENOBARBITONE
BLOOD
Plain tube without gel
4ml
BLOOD
Plain tube without gel
4ml
Gel tube unsuitable. Trough sample.
10-40 mg/L
28 hr
Gel tube unsuitable. Trough sample.
5-20 mg/L
28 hr
PHENYTOIN
3 months
PHOSPHATE
BLOOD
GEL
4ml Gold top tube
0.8-1.5 mmol/L
1 day or 90
min in
emergency
PHOSPHATE
URINE
24hr urine bottle
24hr urine in special
bottle.
Contact Laboratory
for bottle
15.0 - 50 mmol/24hr
1 day
PIPECOLIC ACID
2mL Li heparin blood
(green top tube)
0.5 mL CSF
5 mL urine in plain
container
For interpretation see
report
4-6 weeks
Sent to Referral Laboratory
P a g e | 48
TEST
SAMPLE
REQUIREMENTS
COMMENT
PLACENTAL ALP
Contact Laboratory
Sent to Referral Laboratory
PORPHYRINS
BLOOD
EDTA
4ml
Protect from light.
Send to lab
immediately
Contact Laboratory. All samples are sent
to Referral Laboratory for diagnosis.
Plasma porphyrin
screen
2-3 weeks
PORPHYRINS
URINE
Early Morning Urine
(EMU) in plain
universal bottle.
Protect from light. Send to Lab
immediately
Total porphyrins: <20
nmol/mmol creatinine
24hrs – 3
weeks
POTASSIUM
10ml
FAECES
Plain universal
Request size of stool
sample (5.0g-10g).
Paediatric sample
minimum (0.5g).
BLOOD
GEL
4ml Gold top tube
Expected
TAT
3 months
Porphyrinogen: <1.5
umol/mmol creatinine
To exclude forms of
acute porphyria, the
sample should be
collected during an
acute episode.
PORPHYRINS
REFERENCE
RANGE
Delta-aminolevulate:
<3.8 umol/mmol
creatinine (adult)
<5.2 umol/mmol
creatinine (child)
Protect from light. Send to Lab
immediately.
Total porphyrins:
<200 nmol/g dry
weight
24 hrs
Level increases markedly with haemolysis
or delayed separation.
3.5-5.3 mmol/L
1 day or 90
min in
emergency
P a g e | 49
TEST
SAMPLE
REQUIREMENTS
POTASSIUM OUTPUT
URINE
24 hr urine collection
PREGNANCY TEST
BLOOD
GEL
4ml Gold top tube
PRIMIDONE
REFERENCE
RANGE
Expected
TAT
25-125 mmol/24 hr
1 day
See HCG
Phenobarbitone measured
PRO-BRAIN NATRIURETIC
PEPTIDE (PBNP)
BLOOD
GEL
4ml Gold top tube
PROCOLLAGEN III AMINO
PEPTIDE (PIIINP)
BLOOD
GEL
4ml Gold top tube
BLOOD
GEL
4ml Gold top tube
PROGESTERONE
COMMENT
Contact Laboratory for age/sex matched
reference ranges
PROINSULIN
Contact Laboratory
Sample sent to Referral Laboratory
PROLACTIN
BLOOD
GEL
4ml Gold top tube
Levels up to 800 mU/L can be induced by
stress. Contact Laboratory
<125 ng/L excludes
heart failure as cause
of symptoms
1.7-4.2 ug/L
14 days
Follicular 0.6 – 4.7
nmol/L
ovulation phase:
2.4 – 9.4 nmol/L
luteal : 5.3 – 86
nmol/L
post menopause:
0.3 – 2.5 nmol/L
2 days
Male: 86-324 mU/L
Female: 102-496
mU/L
2 days
P a g e | 50
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
PROSTATIC SPECIFIC
ANTIGEN (PSA)
BLOOD
GEL
4ml Gold top tube
Free/Total PSA ratio automatically tested
when <75 yrs and total PSA 2-10 ng/mL.
Contact Laboratory
Male:
40-49
50-59
60-69
70-79
24 hrs MonFri. If
received after
2pm on
Friday, will be
analysed on
Monday
Please see Appendix 3 for probability of
cancer
0-2.5 ng/mL
0-3.5 ng/mL
0-4.5 ng/mL
0-6.5 ng/mL
PROTEIN CREATININE RATIO
(PCR)
URINE (RANDOM)
Yellow top Monovette
or Sterile Universal
<15.0 mg/mmoL
creatinine
1 day
PROTEIN
ELECTROPHORESIS
Total Protein
Electrophoresis
Plasma CHOLINESTERASE
BLOOD
GEL
4ml Gold top tube
Visual interpretation
7 days
PURINE / PYRIMIDINE
SCREEN
URINE
2-5mls urine
Plain universal
Sample sent to Referral Laboratory
PYRUVATE
Exactly 1ml of blood
in special bottle with
preservative
Contact lab (tel 02890 632148) for tube.
Should be analysed with paired plasma
lactate sample.
RENAL STONE
INVESTIGATION
URINE
2x24hr collections
Contact Laboratory for information
See cholinesterase
3-4 weeks
Contact lab
(tel 02890 633064)
4-6 weeks
24hrs – 4
weeks
P a g e | 51
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
RENIN (Direct)
BLOOD
EDTA
4ml
Send sample to Laboratory immediately (
Please DO NOT send on ice. MUST be
received in lab within 30 minutes
Normal Healthy
Adults: 3.18 - 32.61
pg/mL
Supine:
2.71 - 16.51 pg/mL
Upright:
5.41 - 34.53 pg/mL
2 weeks
RHEUMATOID FACTOR
BLOOD
GEL
4ml Gold top tube
Please note that Rheumatoid Factor for
primary care is no longer available.
Samples will instead be tested for antiCCP antibody which is a more specific test
for rheumatoid arthritis.
0-14 IU/ml
3 days (MonFri) If
received after
2pm on
Friday, will be
analysed on
Monday
SALICYLATE
BLOOD
GEL
4ml Gold top tube
Take sample >4hr after overdose
Antiinflam. 0-30 mg/L
Toxicity >750 mg/L
1 day or 90
min in
emergency
SELENIUM
BLOOD
Plastic heparinised
tube (orange cap) or
blue/black topped
Trace Elements bottle
6mL
BLOOD
GEL
4ml Gold top tube
Special tube – contact laboratory for
supply.
0.60-1.30 umol/L
1 week
Measured in all female patients when
testosterone is requested and used to
calculate Free Androgen Index
FEMALES
20-49yrs: 32.4 – 128
nmol/L
≥50yrs: 27.1 – 128
nmol/L
2 days
SEX HORMONE BINDING
GLOBULIN (SHBG)
P a g e | 52
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
SIALIC ACID
URINE (FRESH)
Plain universal
10ml urine
Sample sent to Referral Laboratory
Qualitative report
4-6 weeks
SIROLIMUS (RAPAMUNE)
BLOOD
EDTA
4ml
Trough Sample
Same day if
received by
12 noon
SODIUM
BLOOD
GEL
4ml Gold top tube
Monotherapy: 12-20
ng/mL
With Cyclosporin:
4-12 ng/Ml
135-145
mmol/L
SODIUM OUTPUT
URINE
24h collection
40 – 220 mmol/24h
1 days
SOMATOSTATIN
BLOOD ON ICE
4ml
EDTA
Transport to lab immediately on ice
0-50 ng/L
2-4 weeks
STEROID PROFILE
URINE
Contact Laboratory
Contact Laboratory
Sample sent to Referral Laboratory
STEROID SULPHATASE
SULPHAEMOGLOBIN
1 day or 90
min in
emergency
Sample sent to Referral Laboratory
< 0.4%
SULPHONYLUREA
BLOOD
Heparinised blood (no
gel)
4ml
Contact Laboratory
Sample sent to Referral Laboratory
SWEAT TEST
SWEAT
Age related
Contact Laboratory to arrange an
appointment for a sweat test
(tel 02890 632148)
1-5 days
P a g e | 53
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
TACROLIMUS, FK 506
BLOOD
EDTA
4ml
Trough sample.
5.0-15.0 ug/L
Same day if
received by
12 noon
TEICOPLANIN
BLOOD
GEL
4ml Gold top tube
BLOOD
GEL
4ml Gold top tube
Contact Microbiology for clinical advice.
See Belfast Trust Guidelines for empirical
antibiotic prescribing in hospitalised adults.
Please see Appendix 4 for Tanner stage
reference ranges
Trough Level 20 –
60mg/L
1 day
Males 20-49yrs
11.4 – 27.9 nmol/L
Females 20-49yrs
0.28 – 1.7 nmol/L.
2 days
TESTOSTERONE
THALLIUM
THALLIUM
THEOPHYLLINE
THIOPENTONE
BLOOD
Blue/black topped
trace metal bottle.
6mL.
URINE
24 hr urine bottle
24 hr urine collection
BLOOD
GEL
4ml Gold top tube
BLOOD
Plain ( no gel)
4mL Red top tube
Male >50 yrs
9.47 – 28.3 nmol/L
Females >50 yrs
0.1-1.39 nmo/L
< 5 nmol/L
2 weeks
Metal Bedpan must NOT be used to
collect urine
< 5 nmol/L
2 weeks
Overdose: Acute >100 mg/L
Chronic >60mg/L may need
haemodialysis. Monitor theophylline and
potassium levels every 2-3 hours.
By special arrangement only.
Contact Laboratory
10-20 mg/L
1 day
Special tube – contact laboratory for
supply.
1 day
P a g e | 54
TEST
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
THIOPURINE METHYL
TRANSFERASE (TPMT)
BLOOD
EDTA
4ml Purple top tube
Sent to Referral Laboratory
14 days
THIOPURINE METABOLITES
BLOOD
EDTA
4Ml Purple top tube
Sent to Referral Laboratory
14 days
THYROGLOBULIN
BLOOD
Gel
4ml Gold top SST
THYROGLOBULIN ANTIBODY
BLOOD
Gel
4ml Gold top SST
Analysed with
thyroglobulin
THYROID PROFILE
Free Thyroxine (fT4)
Thyroid Stimulating Hormone
(TSH)
BLOOD
GEL
4ml Gold top tube
fT4:12-22 pmol/L
TSH: 0.27-4.2mU/L
(adult)
for age and
pregnancy related
reference ranges
please contact lab
1 day
THYROID STIMULATING
HORMONE (TSH)
BLOOD
GEL
4ml Gold top tube
0.27-4.2mU/L (adult)
1 day
1.6-60 ng/ml
for age and
pregnancy related
reference ranges
please contact lab
Thyroglobulin antibody >115 IU/L
invalidates thyroglobulin result.
Expected
TAT
14 days
<40 IU/L
for age and
pregnancy related
reference ranges
please contact lab
P a g e | 55
TEST
SAMPLE
REQUIREMENTS
COMMENT
TOBRAMYCIN
BLOOD
GEL
4ml Gold top tube
Contact Microbiology for clinical advice.
See Belfast Trust Guidelines
for
empirical antibiotic prescribing
in hospitalised adults.
TOTAL PROTEIN
BLOOD
GEL
4ml Gold top tube
60-80 g/L
1 day
TOTAL PROTEIN
URINE
24hr urine collection
bottle
24 hr urine collection
<0.2 g/24hr
1 day
TOTAL PROTEIN
CSF
Plain universal
0.5ml
BLOOD
GEL
4ml Gold top tube
0.15-0.45 g/L
1 day
1.78-2.86 g/L
1 day
Qualitative report
4-6 weeks
Qualitative report
14 days
TRANSFERRIN
TRANSFERRIN GLYOFORM
ANALYSIS
BLOOD
SERUM OR PLASMA
2ml
TRANSFERRIN ISOFORMS
(CARBOHYDRATE-DEFICIENT
TRANSFERRIN)
BLOOD
Contact Laboratory
2ml
Not valid in infants <3 weeks old. Please
allow at least 4-6 weeks after a
transfusion.
Screen for Congenital Disorders of
Glycosylation. Not a suitable test for
alcohol abuse.
Not valid in infants <3 weeks old. Contact
Laboratory
REFERENCE
RANGE
Expected
TAT
1 day
P a g e | 56
TEST
TRICYCLICS SCREEN (See
Drug Screen - Serum)
TRIGLYCERIDE
SAMPLE
REQUIREMENTS
COMMENT
REFERENCE
RANGE
Expected
TAT
BLOOD
GEL
4ml Gold top tube
Patient must fast for 14 hrs before sample
is taken
< 2.2 mmol/L
28 hr
3.1-6.8 pmol/L
1 day
TRIIODOTHYRONINE
(FREE T3)
BLOOD
GEL
4ml Gold top tube
TRIMETHYLAMINE
Contact Laboratory
(tel 02890
632148/633064)
Sample sent to Referral Laboratory
TRIPLE TEST (DOWNS)
Contact Laboratory
Sample sent to Referral Laboratory
TROPONIN T
(HIGH SENSITIVITY)
BLOOD
GEL
4ml Gold top tube
Sample taken at presentation
(time 0)
Second sample at +3h
< 14ng/L
See Appendix 2
For more
information.
90 min in
emergency
TRYPSIN
BLOOD
EDTA ON ICE
4ml Purple top Tube
Transport on ice to Laboratory immediately
120-540 U/L
4 weeks
Male 0.20-0.43
mmol/L
Female 0.14 – 0.36
mmol/L
1.5 - 4.5 mmol/24hr
2-4 weeks
4-6 weeks
TUMOUR MARKERS
See individual tests
URATE
BLOOD
GEL
4ml Gold top tube
URATE
URINE
24 hr collection bottle
24 hr collection
1 day
P a g e | 57
TEST
SAMPLE
REQUIREMENTS
UREA
BLOOD
GEL
4ml Gold top tube
URINE
24 hr collection
BLOOD
GEL
4ml Gold top tube
UREA
VALPROATE
REFERENCE
RANGE
Expected
TAT
2.5-7.8 mmol/L
1 day
430-710 mmol/24 hr
1 day
Poor correlation
between blood levels
and therapeutic
effect.
Trough Level 10 –
15mg/L
1day
0-25 pmol/L
0-120 ng/L
14 days
Sample sent to Referral Laboratory
See report
4-6 weeks
BLOOD
GEL
4ml Gold top tube
BLOOD
Contact Laboratory
BLOOD
GEL
4ml Gold top tube
Protect from light. Analysed with Vitamin E
and beta-carotene
1.1-3.5 umol/L
5-14 days
BLOOD
LITHIUM HEPARIN
ON ICE.
Protect from light.
Send to Laboratory on ice within 30 mins
of taking sample. Protect from light.
VANCOMYCIN
BLOOD
GEL
4ml Gold top tube
VASOACTIVE INTESTINAL
POLYPEPTIDE (VIP)
BLOOD
EDTA ON ICE
4ml Purple top tube
BLOOD
Lithium Heparin
2ml Green top
VERY LONG CHAIN FATTY
ACIDS (VLCFA)
VITAMIN A
VITAMIN B1 (THIAMINE)
VITAMIN B12
VITAMIN C (ASCORBATE)
COMMENT
Trough sample
Contact Microbiology for clinical advice.
See Belfast Trust Guidelines for
empirical antibiotic prescribing in
hospitalised adults.
Transport on ice to Laboratory immediately
Sample sent to Referral Laboratory
1 day
2 weeks
191-663 ng/L
140-190 ng/L
(indeterminate range)
1 day
>32 umol/L Normal
10 – 32 umol/L Risk
of
Vit C deficiency
14 days
P a g e | 58
TEST
SAMPLE
REQUIREMENTS
COMMENT
4ml Green top tube
VITAMIN D
BLOOD
GEL
4ml Gold top tube
Must be received by laboratory within 24hr
of venepuncture
Clinical justification for request MUST be
provided.
REFERENCE
RANGE
< 10 umol/L Severe
risk of Vit C
deficiency
>50 nmol/L
sufficient
Expected
TAT
3 days
30-50 nmol/L
Vit D insufficiency
<30 nmol/L
Vitamin D deficiency
VITAMIN E
BLOOD
GEL 4ml Gold top
tube
Analysed with Vitamin A and betacarotene
WHITE CELL CYSTINE
BLOOD
Lithium Heparin
3ml Green top
Useful in diagnosis of Cystinosis and
monitoring Cystinosis treatment
For urgent delivery. Send Mon-Thurs AM
only (to arrive in lab before 1pm).
WHITE CELL ENZYMES
ZINC
16.0-35.0 umol/L
14 days
8.0 - 15.0 umol/L
1 week
SEE LYSOSOMAL ENZYME SCREEN
BLOOD
Blue/black topped
trace metal bottle.
6mL.
Special tube – contact laboratory for
supply.
Green topped heparinised bottles are
contaminated with zinc and must NOT be
used
P a g e | 59
APPENDICES
The following provide additional information or useful guidance referred from the individual test information or
the laboratory reports.
Appendix 1: Electronic alert systems (e-alerts) for Acute Kidney Injury
Appendix 2: Guidelines on Use of High Sensitivity Troponin T (hsTnT) June 2014
Appendix 3: BHSCT Clinical Biochemistry Guidelines Free Prostate Specific Antigen (fPSA)
Appendix 4: Puberty related Reference Ranges for Testosterone
Appendix 5: BHSCT Clinical Biochemistry Guidelines Alpha – Feto Protein (AFP) Reference Ranges
Appendix 6: Laboratory Investigation of Pheochromocytoma and Paragangliomas (PPGLs)
P a g e | 60
Appendix 1: Electronic alert systems (e-alerts) for Acute Kidney Injury GAIN Guidelines 2014
P a g e | 61
Appendix 1: Electronic alert systems (e-alerts) for Acute Kidney Alert
Electronic alert systems (e-alerts) for Acute Kidney Injury
Acute kidney injury (AKI) is common, often preventable and associated with high morbidity and
mortality1,3. It is diagnosed by a reduction in urine output or a rise in serum creatinine from a
baseline value1,2.
Delay in diagnosing AKI or failure to recognize AKI significantly contributes to the poor outcomes of
patients with AKI3. There is therefore an urgent need to improve early recognition of AKI3.4.
The development of electronic alert systems (e-alerts) for AKI would seem to be an effective way
of doing this4. The electronic alert system uses laboratory data to ‘flag’ or ‘alert’ when a patient
has a change in serum creatinine level, which may identify that patient as having AKI. This ‘alert’
will appear on the screen when the creatinine result is looked up. The responsibility for looking up
the result and taking the appropriate clinical action remains with the clinician(s) responsible for the
patient.
The electronic alert systems are not designed to replace timely clinical investigation, examination
and management of patients but are an additional ‘warning system’.
It is hoped that electronic alert systems for AKI combined with better identification of the ‘at risk’
patient and improved clinical assessment will trigger earlier recognition and management of
persons with AKI. This should improve patient outcomes.
References:
1. NI Guidelines for Acute Kidney Injury (2010) http://www.gainni.org/images/Uploads/Guidelines/GAIN%20-%20Acute%20Kidney%20Injury%20Guideline.pdf
2. KDIGO Clinical Practice Guideline for Acute Kidney Injury. Kidney Disease: Improving Global
Outcomes (KDIGO) Acute Kidney Injury Work Group. KDIGO Clinical Practice Guideline for Acute
Kidney Injury. Kidney inter., Suppl. 2012; 2: 1–138.
3. NCEPOD Acute Kidney Injury: Adding Insult to Injury (2009)
4. RCPE UK Consensus Conference on “Management of acute kidney injury: the role of fluids, e-alerts
and biomarkers” 16 & 17 November 2012
P a g e | 62
Appendix 2: Guidelines on Use of High Sensitivity Troponin T
(hsTnT) June 2014
Key Messages:
1. Clinical context is critical to the interpretation of any hsTnT result.
2. All patients under investigation for possible acute coronary syndrome (ACS)
should have hsTnT measured at presentation (Time 0 )
3. The 99th percentile for hsTnT in a healthy population is 14 ng/L.
4. The lower reporting limit for the hsTnT assay is 3 ng/L
The range 3-14 represents ≈ 50 % of the distribution in the healthy population
5. An initial hsTnT result < 3 ng/L at presentation has a high negative predictive
value for ACS, if several hours post ictus. However, this may not hold for early for
early presenters.
6. The higher the initial hsTnT (above 14 ng/L) ,with evidence of myocardial
ischemia, the more likely the diagnosis of AMI.
7. When further investigation is indicated a second sample at + 3 hours after the
initial hsTnT sample is recommended. The relative change between the 0 and 3
hour sample can help identify acute myocardial necrosis.
8. A change of >20% in 3h with one value >14 ng/L is consistent with a diagnosis
of an ACS *Universal definition of MI, 2012)
9. The higher the delta change (%), the more likely the diagnosis of MI and a
change >100% has a high positive predictive value.
10. Results produced using the laboratory hsTnT assay are not directly comparable
to any other troponin assay result.
P a g e | 63
Appendix 2: Guidelines on Use of High Sensitivity Troponin T
(hsTnT)
Interpretation of hsTnT results:
Presentation – 1st sample (Time 0)
hsTnT: < 3ng/L
On presentation this result has a high negative predictive value for an ACS. However, an early
or rapid presentation may require a second sample at +3Hrs.
hsTnT: 3-14 ng/L
By definition approximately 50 % of the asymptomatic population may have a hsTnT in this
range. A second sample at +3Hrs may be required to rule AMI/ACS in or out, in the appropriate
clinical context.
hsTnT: ≥ 14ng/L
A result ≥ 14ng/L is outside the population reference range and may be due to acute or nonacute disease. A second sample collected at +3Hrs can assist diagnosis where the ECG is
inconclusive.
2nd Sample at +3Hrs
The relative % change in the hsTnT between these two samples can be used to support the
diagnosis of ACS or alternatively assist the rule-out of acute myocardial necrosis
 If the second hsTnT remains < 14ng/l this does not meet the criteria for the universal
definition of MI (Definition requires hsTnT > 99th percentile [14 ng/L])
 A hsTnT change up to 20 % can be explained by normal sources of variation and so a
change ≤20% has a high NPV for rule-out of acute myocardial necrosis
 A hsTnT > 14ng/L and a change > 20% is considered significant and, alongside
evidence of cardiac ischemia, supports a diagnosis of AMI. The greater the rise over
3Hrs the more likely a diagnosis of ACS and a change >100% approaches 100% PPV.
 In some cases a 3rd sample at 6Hrs post presentation may help to clarify the clinical
picture.
P a g e | 64
Appendix 3: BHSCT Clinical Biochemistry Guidelines Free Prostate
Specific Antigen (fPSA)
Prostate specific antigen (PSA) exists in serum in both bound and free forms. The free form
(Free PSA [fPSA]) includes enzymatically inactive pre-PSA, pro-PSA, clipped-PSA and the
enzymatically active form of free PSA. The total PSA (TPSA) in the circulation corresponds to
the sum of circulating free PSA (fPSA) and PSA bound as a stable complex to alpha-1antichymotrypsin (PSA-ACT). The free fraction constitutes from 5% to more than 40% of the
total. Calculation of percentage free PSA (%fPSA = (fPSA / TPSA)x100) is recommended by
the National Academy of Clinical Biochemistry (NACB) and the European Group on Tumour
Markers (EGTM) as an aid distinguishing men with prostate cancer from men with benign
disease in selected high-risk groups, especially when total PSA values are between 4 and 10
ng/L. The use of % fPSA can enhance the specificity of TPSA in detecting prostate cancer
and reduce the number of unnecessary biopsies.
% fPSA decision limits
There is no definitive reference limit for %fPSA and results should be reviewed in line with
clinical presentation. There is a higher probability of cancer when the %fPSA is lower (See
Table 1).
%fPSA
% Cancer Probability
0-10
56 %
10-15
28 %
15-20
20 %
20-25
16 %
PLEASE NOTE: THE FOLLOWING CAN EFFECT PSA (Total & Free) LEVELS
- BPH, acute and chronic prostatitis, UTI, urinary retention cause elevations
- TURP & Needle biopsy cause significant elevations (Wait 6 weeks before
sampling)
- Prostate massage and ejaculation (possible transient elevations)
- Drugs: Finasteride and Dutasteride, 5-alpha-reductase inhibitors used to treat
BPH reduce PSA levels by approx. 50%.
References:
Association of Clinical Biochemists in Ireland: Guidelines for the use of tumour markers. 4 th
ed. 2010. http://www.acbi.ie/Article.asp?pID=231.
The National Academy of Clinical Biochemistry: Use of tumour markers in testicular, prostate,
colorectal, breast and ovarian cancers. 2009.
http://www.aacc.org/members/nacb/LMPG/OnlineGuide/PublishedGuidelines/major/Documen
ts/TumorMarkers.pdf
P a g e | 65
Appendix 4 Puberty related Reference Ranges for Testosterone
The following table shows the expected ranges for testosterone in a healthy population1.
Tanner stage was characterised according to the method of Marshall and Tanner 2,3.
Testosterone nmol/L
Tanner Stage
Male (age7-18yrs)
Female (age 8-18 yrs)
1
<0.8
<0.4
2
<7.3
<0.8
3
0.6-23.4
0.2-1.4
4
4.3-29.0
0.4-1.8
5
7.6-27.0
0.5-2.0
References
1. Testosterone II Kit Insert 2012-05, V 6.0. Roche Diagnostics.
2. Marshall WA, Tanner JM. Variations in the pattern of pubertal changes in boys. Arch
Dis Childh 1970;45:13-23.
3. Marshall WA, Tanner JM. Variations in the pattern of pubertal changes in girls. Arch
Dis Childh 1970;45:291-303.
P a g e | 66
Appendix 5
BHSCT Clinical Biochemistry Guidelines
Alpha – Feto Protein (AFP) Reference Ranges
In utero, the AFP serum concentration increases until it reaches a peak at 14
weeks gestation and then decreases steadily. AFP synthesis almost ceases
at parturition resulting in an exponential fall of AFP serum concentrations in
the first year of life. The age-related reference range is shown below:
Age
AFP Reference Range (kU/L)
Birth
50,000 – 150,000
2 weeks
7,000 – 20,000
4 weeks
1,500 – 2,500
6 weeks
200 - 400
8 weeks
50 - 100
10 weeks
6 - 12
3 months to 50 years
3 - 10
50 – 70 years
< 15
70 – 90 years
< 20
NOTE: Higher values are seen in the premature infant and the degree
of immaturity will influence the age (from birth) at which basal levels
are attained
th
[Reference Ranges from PRU handbook of Clinical Immunochemistry – 9 Edition]
P a g e | 67
Appendix 5
BHSCT Clinical Biochemistry Guidelines
Alpha – Feto Protein (AFP) Reference Ranges
Pregnancy-related Ranges
AFP is detectable in maternal serum and in amniotic fluid during pregnancy. AFP concentrations peak
around 30 weeks gestation and fall to normal non-pregnant concentrations at delivery. Reference ranges
for maternal serum at different gestational ages (in weeks), are shown below:
AFP Reference Range (kU/L)
th
Weeks gestation
50 centile
th
10 centile
th
95 centile
(Median)
Maternal
Serum
14
< 10
19
50
15
< 10
24
60
16
10
31
69
17
11
37
81
18
13
43
98
19
15
50
115
20
18
60
123
21
21
67
142
th
[Reference Ranges from PRU handbook of Clinical Immunochemistry – 9 Edition]
AFP as a Tumour Marker
AFP concentrations are elevated in 70-95% of patients with primary hepatocellular carcinoma and in
patients with non-seminomatous germ cell tumours (NSGCT).
AFP Reference Range (tumour marker):
0 - 10 kU/L
References
th
Protein Reference Units - Handbook of Clinical Immunochemistry 9 Edition (2007)
AFP Kit Insert – Roche Cobas Modular Analytics (2012)
P a g e | 68
Appendix 6 Laboratory Investigation of Pheochromocytoma and
Paragangliomas (PPGL)
PPGLs are potentially lethal yet surgically curable causes of endocrine Hypertension.
They are rare and clinical presentation is varied with an often insidious onset.
However, once clinical suspicion is aroused it is imperative that reliable laboratory
investigations are employed to identify the tumours.
Urine fractionated metanephrines are now recognised as the best first line
investigation. Metanephrine, Normetanephrine and 3 Methoxytyramine are natural
metabolites of the catecholamines Adrenaline, Noradrenaline and Dopamine
respectively and measurement of excretion of the fractionated metanephrines in urine
over 24 hours now replaces testing of the free catecholamines.
Fractionated Metanephrine excretion within the reference interval reliably excludes a
PPGL where the clinical suspicion arises due to hypertension and associated
symptoms (NPV >97%). This requires only a single 24 h urine collection as opposed to
the X 3 collections recommended for traditional free catecholamine testing. A small
number of false positive results is to be expected and any patients with an abnormal
results should be referred for follow up investigations including imaging when this is
warranted.
Sample requirements
Patients should be asked to avoid stimulants (eg coffee) and paracetamol for the day
before starting a 24h urine collection. It is not necessary to stop any other medication
which could cause a false positive result however, this may be necessary if an
abnormal result is obtained. Please provide details of current medication on the
request form.
Collection requires a special 24h urine bottle containing preservative (same as for
catecholamine testing). It is most convenient for the patient to void the bladder on first
rising in the morning (discard), note date and time on label/form; collect all urine
voided for the next 24h including final void at the same time the following morning
(note date and time again). Transport to lab without delay.
Reference values: 24h Urine Collection
Normetanephrine Output 440 – 2960 nmol/24 hours
Metanephrine Output
141 – 1289 nmol/24 hours
Reference
Pheochromocytoma and Paraganglioma: An Endocrine Society Clinical Practice
Guideline. Lenders et al, J Clin Endocrinol Metab 99 (6), 1915-1942: 2014.
P a g e | 69
Department of Haematology
Laboratory Services
The Department of Haematology provides a comprehensive routine and specialist
regional diagnostic, interpretative and clinical advice service. The repertoire of
investigations and required specimens are outlined in this document. Each test
request accepted by the Haematology laboratory service shall be considered as an
agreement; which will take into account the request, the examination and the report.
General Haematology and Blood Transfusion services are provided at The Royal
Hospitals, Belfast City Hospital and the Mater Hospital. The specialist regional
services are located at the Belfast City Hospital laboratory.
Opening times to the laboratories:
 The Royal Hospitals & Belfast City Hospital
Monday to Friday
9:00am - 5:15pm
Sat, Sun & Bank Holidays
9:00am - 5:00pm
Emergency Out of Hours Service:
Monday to Friday
5:00pm - 9.00am
Sat, Sun & Bank Holidays
5:00pm - 9.00am
 The Mater Hospital
Opening hours of the Haematology and Blood Transfusion Laboratory:
Monday to Friday
8:30am – 4:00pm
Sat, Sun & Bank Holidays
8:30am - 12:00 midday
P a g e | 70
Outside of these hours an emergency Out of Hours Service is provided by The
Royal Hospitals.
During these hours Clinical staff should contact:


RVH Blood Bank 028 9063 3663 or 028 9063 2359
RVH Haematology Laboratory 028 9063 3669 or 028 90632526
* The Out of Hours service must only be used for urgent requests which may
influence immediate patient management.*
The out of hours repertoire is restricted to CBC, coagulation screening, INR, APTT
for monitoring of unfractionated heparin, D-dimers in specific circumstances, ESR for
?temporal arteritis, blood grouping and provision of blood and blood products. Other
investigations which may be required, for example over long weekends and public
holidays can be arranged as appropriate by prior discussion with the duty Consultant
Haematologist.
Clinical Advice and Interpretation: Where appropriate, interpretative information
will be reported with numerical results. For more detailed interpretation or clinical
advice please contact the duty laboratory Specialist Registrar (SpR) via switchboard
at the Belfast City Hospital (BCH) or the appropriate consultant.
There is a 24-hour consultant-led medical on-call service available for interpretative
advice and clinical consultation in urgent cases, including acceptance of patients
who are found to have primary haematological disorders. The duty SpR and/or
consultant can be contacted via BCH switchboard or via Ward 10 North, BCH.
……………………………………………………………………………….
P a g e | 71
Location of Haematology Laboratories within the Belfast Trust
Belfast City Hospital Laboratory
C Floor, BCH Tower
Lisburn Road, Belfast,
BT9 7AB
Tel. 028 90 329241
Royal Victoria Hospital Laboratory
Kelvin Building, RVH
Grosvenor Road, Belfast,
BT12 6AB
Tel. 028 90 240503
Royal Belfast Hospital for Sick Children
Grosvenor Road, Belfast,
BT12 6AB
Tel. 028 90 240503
Mater Hospital Laboratory
Crumlin Road, Belfast,
BT14 6AB
Tel. 028 90 741211
P a g e | 72
HAEMATOLOGY, BLOOD TRANSFUSION & STEM CELL BANK
The Royal Hospital Laboratory, Kelvin Building, Grosvenor Road, Belfast, BT12 6AB
Belfast City Hospital, C Floor, BCH Tower, Lisburn Road, Belfast , BT9 7AB
Mater Hospital, Crumlin Road, Belfast BT14 6AB
General Enquiries
RVH
028 9063 3663
Blood Bank
/ 2359
BCH
MIH
028 950 40987
/ 40988
028 950 41332/
41328
Routine Haematology
028 9063 3669
028 950 40922
/ 40921
028 950 41508/
41328
Coagulation
028 9063 2526
028 950 40921
028 950 41508/
41328
Paediatric Haematology
028 9063 2098
Haemostasis & Thrombosis
028 950 40910
Haemato-Oncology (Flow Cytometry)
028 950 40913
Stem Cell Bank
028 950 40912
Andrology
028 950 40911
Red Cell Investigations
028 950 40915
Secretaries office
028 950 47989
028 9080 2223
Bleep 1702
Contact Royal
Hospitals,
Blood Bank
Out of Hours Emergency
028 9063 3663
028 9063 3663
On-call Registrar &/or Consultant
BCH
switchboard/
028 9032 9241
BCH
switchboard/
028 9032 9241
BCH
switchboard/
028 9032 9241
P a g e | 73
Professional Contacts
Office: 028 950 48002
Clinical Lead
Dr RJG Cuthbert
Lab: 028 950 40913
Quality Manager
Mrs Alison Geddis
028 906 32552
Laboratory Manager – Blood Transfusion &
Stem Cell Bank
Mrs Audrey Savage
028 906 34021
Laboratory Manager – Haematology
Ms Annette Hobson
028 950 47908
Operational Manager – Blood Bank RVH &
MIH
Mr Matt Gillespie
028 906 33663 /
32531
Operational Manager – Blood Bank BCH
Post Vacant
028 950 40990
Mrs Robyn McConnell
028 950 45343
Miss Joy Gallagher
028 906 38298
Operational Manager – Stem Cell Bank
Ms Liz Moody
028 950 40912
Operational Manager – Specialist
Haematology
Mr Gary Beattie
028 950 40913
Molecular Haemato-Oncology
Dr Mark Catherwood
Operational Managers – Haematology
Office: 028 950 48138
Lab:
028 950 40914
Office: 028 950 47746
Molecular Red Cell Disorders
Dr Melanie Percy
Lab:
028 950 40914
Office: 028 950 48062
Molecular Haemostasis
Dr Paul Winter
Lab:
Laboratory Registrar
028 950 40914
Rotation
028 950 40911/40913
Dr Mary Drake
028 950 48141
Dr Paul Kettle
028 950 47974
Dr Michael Quinn
028 950 47831
Dr Oonagh Sheehy
028 950 48069
Lymphoma/Myeloma Service:
P a g e | 74
Dr Claire Arnold
028 950 48036
Dr Damian Finnegan
028 950 47940
Dr RJG Cuthbert
028 950 48002
Prof MF McMullin
028 950 48008
Haemostasis & Thrombosis Service
Dr Gary Benson
028 950 47977
Paediatric Haematology Service
Dr Christine Macartney
028 906 33632
Regional Haemovigilance Co-ordinator
Ms Aine McCartney
028 906 34078
Leukaemia/BMT Service:
P a g e | 75
Instructions for completion of Haematology request forms
Attention to detail is essential to ensure that the right result is sent out on the right
patient. Specimens will not be accepted for analysis where the following essential
criterion in the Minimum Data Set is not met on the form or the specimen container.
Sample
Essential
Desirable
Patients first name*
Date and time of specimen
Surname*
Destination for report
Date of birth and/or
Request Form
Hospital number (or H&C
number)
Sample PID labels are
preferred – except for
Blood Transfusion.
Patients first name*
Clinical details
Surname*
Patient’s address
Date of birth and/or hospital
number (or H&C number)
Gender
Time of specimen
Patient’s location and
destination for report
Patient’s consultant or GP
Test request
Name of requesting
practitioner
Date of specimen
* or proper coded identifier
Hospital or GP practice
addressograph labels are
preferred.
Additional investigations: Telephone the lab within 4 hours to request additional
investigations on a specimen, for example reticulocyte count, blood film, red cell
investigations, coagulation tests etc.
P a g e | 76
Instructions for completion of Blood Transfusion request forms
Requests for blood grouping &/or ordering blood products must be completed by a
doctor or designated nurse. The collection and labelling of specimens for blood
transfusion may be delegated to designated staff who have had the appropriate
training. The laboratory will undertake testing and issue of blood/blood products only
on receipt of a legible request form and a correctly labelled specimen. Incorrectly
completed forms or specimens will be rejected according to national guidelines.
Essential
Sample
Desirable
First name
Surname
Date of birth
Hospital number/H&C
number
Signature
Date of specimen
Patient details MUST be
hand-written on the
specimen bottle.
Request Form
First name
Patient’s address
Surname
Consultant
Date of birth
Include clinical details:
Gender
Blood group (if known)
Hospital number/H&C
number
Previous transfusions
Signature
Date/Time of specimen
Obstetric history
-
Any known red cell antibodies
Previous transfusion reactions
Any recent anti-D administration
Test request
Product requirement &
date/time required
An addressograph label may be used
on the request form as long as the
Ward/Consultant and Date/Time are
recorded.
P a g e | 77
It is the responsibility of the staff taking the specimen to complete the patient details
on the specimen tube immediately after taking the specimen.
An addressograph label must not be used on the specimen. Patient details
must be confirmed against the patient’s wrist-band, and by direct questioning of the
patient according to Trust policy (see hyperlink below).
BHSCT Policy on the Administration and Management of blood components:
http://intranet.belfasttrust.local/policies/Documents/Blood
components
Administration and management of the transfused patient in the hospital setting.pdf
Pre-labelling of specimens is highly dangerous and is not
permitted.
The section on the request form labelled “I confirm that the patient transfusion history
is correct and the patient identification details correspond to the details of the patient
and the specimen tube” must be signed by the person taking the blood sample.
Additional Transfusion Requests: The sample will be held for 7 days. If the
patient has been transfused more than 72 hours previously, a new sample is
required according to the following guidelines:

Patient transfused within:
New sample to be taken:
3 days - 28 days
72 hours before transfusion
28 days - 3 months
1 week before transfusion
Irregular Antibodies:
Please note that if a patient is found to have a red cell alloantibody or a positive
Direct Antiglobulin Test (Coomb’s test) it will take longer to select compatible blood.
Please contact the blood bank to discuss emergency cases or pre-operative cases
for whom blood is required in theatre. Patients in the latter category should not be
sent to theatre until you have confirmed that compatible blood is available.
P a g e | 78
Unconscious /unidentified patients: A special protocol is applicable to unidentified
patients in A&E and theatres.

Paediatric Transfusion:
(i) Neonates & infants <4 months: An initial sample must be taken from infant &
mother. When infant/mother incompatibility exists the blood will be crossmatched against the mother's plasma. The mother's blood group, antibody
screen and NIBTS antenatal reference number must be sent to the lab.
(ii) Older infants & children: Send only the child’s sample, unless contacted by the
lab.

Blood Tracking and Cold Chain:
Blood/blood product tracking and maintenance of the “cold chain” are mandatory
national requirements of the Blood Safety and Quality Regulations. Blood must be
stored only in designated blood refrigerators and not in any other ward refrigerators.
Blood will be returned to stock 48 hours after the date/time requested as stated on
the request form unless the blood bank is notified that it is still required.
Availability of Results
Turnaround Times



Routine CBC and coagulation screen results are available within 4 hours.
Reports are returned within two days.
Samples marked 'Urgent' are given priority and results are available in <2
hours of receipt of sample.
More specialised tests are batched and results are available two working days
after completion.
Request
Expected TAT’s
Emergency
<2hrs
Routine
<4hrs
Specialist (these include Haemostasis, Red Cell
Investigations, Haemato-Oncology)
2-3 weeks
Molecular (these include Haemostasis, Red Cell
Investigations, Haemato-Oncology)
2-6 weeks
P a g e | 79
Hospital users have access to LabCentre via ward computers. Routine CBC &
coagulation screen results, which have arrived in the lab. before 3.00 pm, will be
displayed by that evening. If further tests, such as a manual differential white cell
counts are required, the results will not be displayed until they are validated. The
accession no. will be displayed, and by telephoning the lab, a provisional result can
be given. For an extremely urgent sample contact the laboratory to have it
prioritised.
LabCentre log-ins for clinicians do not allow access to unvalidated results. Users
must not take clinical decisions based on unvalidated results.
All laboratory results reported are based on professional judgements made by
personnel who have the appropriate qualifications for their job role.
All personnel will have the applicable theoretical and practical background
experience to perform the assigned managerial and technical tasks in accordance
with national, regional and local regulations and professional guidelines.

Health and Safety
All specimens must be treated as a potential hazard.
Specimens of blood, serum and other body fluids from suspected
carriers of Category 3 pathogens (hepatitis B or C and HIV) must be
clearly marked with hazard stickers and enclosed in a sealed
plastic bag. Request forms should also have a hazard sticker.

Laboratory Request Forms with Attached Plastic Specimen Envelope
These specially designed once-only laboratory form/specimen carriers are practical
and easy to use. All blood samples must be transported in these bags and in
accordance with the BHSCT Laboratories Transport Policy.
P a g e | 80
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
BLOOD BANK
ANTIBODY
INVESTIGATION
BLOOD GROUP &
ANTIBODY SCREEN
COLD AGGLUTININ
TITRE
CELL GROUP & DAT
(< 4 MONTHS OLD)
* CROSS-MATCH /
ELECTRONIC ISSUE
DIRECT
ANTIGLOBULIN TEST
(COOMB’S TEST)
KLEIHAUER TEST
Sample sent to Regional
Referral Laboratory,
NIBTS. Contact
laboratory if blood is
required urgently. See
Guidelines in appendix 9.
24 -72hrs
Dependent
on NIBTS
Laboratory
Service.
Blood
EDTA
12mls
Blood
EDTA
6mls
< 4hrs
Blood
EDTA
6mls
24 hrs
Blood
EDTA
2.5mls
< 4hrs
Blood
EDTA
6mls
< 4hrs
Blood
EDTA
4 or 6mls
< 4hrs
Blood
EDTA
6mls
48 hrs
* In an emergency please contact Blood Bank to prioritise sample (TAT < 2hrs)
P a g e | 81
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
N/A
By arrangement with
laboratory staff.
See guidelines in
Appendix 3.
< 4hrs
See individual
parameters
below
See individual
parameters below
< 4hrs or
<2hrs in
emergency
HAEMATOLOGY
BLOOD FILM
COMPLETE BLOOD
COUNT (CBC)
(Causasians)
Blood
Blood
EDTA
EDTA
4mls
4mls
CBC Sample must be
analysed <16 hours from
the bleed time
Neutrophils
Cord – 3rd day
1.5 – 7.0 x
109/L
DIFFERENTIAL WHITE
CELL COUNT (DWCC)
Blood
EDTA
4mls
4th day – 4
years
2.0 – 6.0 x
109/L
Adults
2.0 - 7.5 x 109/L
See guidelines in
Appendix 3.
< 4hrs
P a g e | 82
Lymphocytes
Cord – 3rd day
2.0 – 5.0 x
109/L
4th day – 4
years
5.8 – 8.5 x
109/L
Adults
1.0 - 3.5 x 109/L
Monocytes
Cord – 3rd day
0.3 – 1.1 x
109/L
4th day – 4
years
0.7 – 1.5 x
109/L
Adults
0.2 - 0.8 x 109/L
P a g e | 83
Eosinophils
Cord – 3rd day
0.2 – 2.0 x
109/L
4th day – 4
years
0.3 – 0.8 x
109/L
Adults
0.04 - 0.4
x109/L
Basophils
Cord – 3rd day
<0.1 x 109/L
4th day – 4
years
<0.1 x 109/L
Adults
0.01 - 0.1
x109/L
P a g e | 84
TEST
SAMPLE
SAMPLE
TUBE
ERYTHROPOIETIN
Blood
Serum
VOLUME
6mls
PRECAUTIONS
REFERENCE
RANGE
2.5 –
10.5mlU/mL
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
2-3 weeks
Maximum in1st
hr (mm):
ESR
Blood
EDTA
4mls
Insufficient samples
(<1.5ml) will be rejected
Male
17 - 50 yrs
10mm
51 – 60 yrs
12mm
61 – 70 yrs
14mm
> 70yrs 30mm
Female
17 - 50 yrs
12mm
51 - 60 yrs
19mm
61 - 70 yrs
20mm
> 70 yrs 35mm
< 4hrs
P a g e | 85
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
G6PD
Blood
EDTA
4mls
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
5.2 –
22.8IU/gHb
By arrangement with
laboratory staff
5 days
Cord
0.54 – 0.68
3 months
0.30 – 0.36
HAEMATOCRIT
(HCT)
Blood
EDTA
4mls
3 – 6 years
0.36 – 0.44
< 4hrs
Male
0.40 - 0.54
Female
0.37 - 0.47
HAEMOGLOBIN
(HB)
Cord
Blood
EDTA
4mls
168 – 212g/L
< 4hrs
P a g e | 86
3rd Day
153 – 221g/L
1st week
150 – 196g/L
3 months
104 – 122g/L
3 – 6 years
120 – 140g/L
Male
130 - 180g/L
Female
115 - 165g/L
P a g e | 87
TAT
By arrangement with
laboratory staff
2-3 weeks
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
HB
ELECTROPHORESIS
Blood
EDTA
4mls
Hb F
Blood
EDTA
4mls
0 – 4%
2-3 weeks
HbA1C
Blood
EDTA
4mls
<53mmol/mol
5 days
HbA2
Blood
EDTA
4mls
1.5 – 5%
2-3 weeks
MALARIAL PARASITES
Blood
EDTA
4mls
PRECAUTIONS
Optimum time for
collection during or just
after a temperature
spike.
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
Sample must be
received in RVH lab.
< 2hrs old
< 4hrs
P a g e | 88
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Cord
32 – 36pg
3 months
MEAN CELL
HAEMOGLOBIN
29 – 33pg
Blood
EDTA
4mls
< 4hrs
3 – 6 years
(MCH)
24 – 30 pg
Adult
27.0 - 32.0 pg
Cord – 6 years
MEAN CELL
HAEMOGLOBIN
CONCENTRATION
(MCHC)
300 - 360 g/L
Blood
EDTA
4mls
< 4hrs
Adult
320 - 360 g/L
Cord
MEAN CELL VOLUME
(MCV)
Blood
EDTA
4mls
110 – 128fL
< 4hrs
P a g e | 89
3rd Day
111 – 121fL
1st week
93 – 131fL
3 months
80 – 96fL
3 – 6 years
73 – 89fL
Adult
76 - 100 fL
P a g e | 90
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PLASMA
HAEMOGLOBIN
Blood
EDTA
4mls
PLASMA VISCOSITY
Blood
EDTA
4mls
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
< 1mg/100ul
1.50 – 1.72 cp
TAT
1 week
Sample sent to
referral laboratory
The Ulster Hospital
1 week
Cord – 2nd
week 120 –
380 x109/L
PLATELET COUNT
Blood
EDTA
4mls
< 4hrs
nd
2 week Adult 150 –
450 x109/L
P a g e | 91
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PYRUVATE KINASE
Blood
EDTA
4mls
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
6.0 – 19.2
IU/gHb
By arrangement with
laboratory staff
1 week
Cord
4.4 – 5.8 x
1012/L
3 months
3.4 – 4.0 x
1012/L
RED CELL COUNT
(RBC)
Blood
EDTA
4mls
3 -6 years
4.1 – 5.5 x
1012/L
Male
4.5 - 6.5 x
1012/L
Female
3.8 - 5.8 x
1012/L
< 4hrs
P a g e | 92
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Sample must be <24
hours old from the
bleed time.
< 4hrs
Qualitative
report
Contact lab if
required urgently
< 4hrs
Quantitative
report
See Guidelines in
Appendix 1
2 -3
weeks
REFERENCE
RANGE
Cord
2-5% (88 - 290
x109/L)
3 months
RETICULOCYTE
COUNT
Blood
EDTA
4mls
0.2-2.0% (6.8 80 x 109/L)
Adults
0.2 -2.5%
(50 – 100
x109/L)
SICKLE CELL TEST
Blood
EDTA
4mls
P a g e | 93
Cord – 3rd Day
9 – 30 x109/L
4th day – 4th
year
WHITE BLOOD CELL
COUNT (WBC)
5 – 20 x109/L
Blood
EDTA
4mls
< 4hrs
4 -7 years
5 - 15 x109/L
Adults
4.0 - 10.0
x109/L
P a g e | 94
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
See “Directions for
obtaining a specimen for
post vasectomy
investigation”
Referrals from patients with
Category 3 status should be
made to the Regional
Infertility Centre.
POST VASECTOMY
Semen
Contact
Andrology
lab, BCH
Ejaculate x1
By arrangement with
Andrology lab, BCH.
Please indicate on the referral
form if the report should be
issued to a Clinician other
than the referring Clinician.
See guidelines in
Appendix 12 for
obtaining a specimen
2-3
weeks
Clinical advice and
interpretation is available –
Please contact Dr Robert
Cuthbert (see contact details)
who will arrange this through
the clinical infertility service.
URINARY MYOGLOBIN
Urine
Sterile
Container
10 – 15mls
Not detected
< 4hrs
P a g e | 95
COAGULATION
TEST
ADAMTS13
SAMPLE
Blood
SAMPLE
TUBE
Citrate
VOLUME
2.7mls
PRECAUTIONS
Fill sample tube to line
indicated on bottle
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
40 – 120%
Contact lab if required
urgently. Sample must
arrive at the
Haematology
Laboratory, BCH within
4 hours of sample being
taken or plasma can be
received frozen.
2-3 weeks
or <6hrs in
emergency
Prophylaxis
ANTI- XA
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.2 – 0.4 iu/ml
< 4hrs
Treatment
0.5 – 1.0 iu/ml
ANTI-THROMBIN III
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
APC RESISTANCE
RATIO
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
(APC-R)
0.8 – 1.2 iu/ml
Also available as part of
Thrombophilia Screen
2-3 weeks
or < 4hrs in
emergency
> 2.20
Also available as part of
Thrombophilia Screen
2-3 weeks
P a g e | 96
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Sample must be
analysed within 4 hours
of the bleed time.
23.4 – 32.4 secs
Also available as part of
Coagulation Screen.
Underfilled,
haemolysed, lipaemic
sample will be rejected.
< 4hrs
PRECAUTIONS
Fill sample tube to line
indicated on bottle
APTT
Blood
Citrate
2.7mls
COAGULATION
FACTOR ASSAYS
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
See individual
analytes
COAGULATION
SCREEN
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
See individual
analytes
D-DIMERS
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
< 0.5g/ml
FACTOR INHIBITOR
ASSAYS
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
FACTOR II
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
2-3 weeks
Underfilled,
haemolysed, lipaemic
sample will be rejected.
Underfilled,
haemolysed, lipaemic
sample will be rejected.
By arrangement with
laboratory staff – see
individual factors
< 4hrs
< 4hrs
2-3 weeks
2-3 weeks
P a g e | 97
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
FACTOR V
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
FACTOR V LEIDEN
Blood
EDTA
4mls
FACTOR VII
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
2-3 weeks
FACTOR VIII
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
< 4hrs
FACTOR IX
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
< 4hrs
FACTOR X
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
2-3 weeks
FACTOR XI
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
< 4hrs
FACTOR XII
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.6 – 1.3iu/ml
< 4hrs
TAT
2-3 weeks
Also available as part of
Thrombophilia Screen
2-6 weeks
P a g e | 98
TEST
SAMPLE
SAMPLE TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
1.9 – 4.0g/l
Also available as part of
Coagulation Screen.
Underfilled, haemolysed,
lipaemic sample will be
rejected.
< 4hrs
Fill sample tube to line
indicated on bottle.
FIBRINOGEN
Blood
Citrate
2.7mls
HITT ASSAY
Blood
Serum
4 or 6mls
Sample must be
analysed within 4 hours
of the bleed time.
Not detected
Fill sample tube to line
indicated on bottle.
INR
Blood
Citrate
2.7mls
LUPUS
ANTICOAGULANT
Blood
Citrate
2 x 2.7mls
Sample must be
analysed within 4 hours
of the bleed time.
Fill sample tube to line
indicated on bottle
< 72hrs
Therapeutic
target range
dependent on
clinical
indication
Underfilled, haemolysed,
lipaemic sample will be
rejected.
< 4hrs
Negative
Also available as part of
Thrombophilia Screen
2-3
weeks
By arrangement with
laboratory staff
< 2hrs
Collagen/ADP
71 – 108 secs
PFA-100
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
Collagen/
Epinephrine
94 – 193 secs
P a g e | 99
TEST
SAMPLE
SAMPLE TUBE
VOLUME
PLATELET
AGGREGATION
Blood
Citrate
6 x 2.7mls
PROTEIN C
Blood
Citrate
2.7mls
PRECAUTIONS
Fill sample tube to line
indicated on bottle
REFERENCE
RANGE
0.7 – 1.4 iu/ml
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
By arrangement with
laboratory staff
< 4hrs
Also available as part of
Thrombophilia Screen
2-3
weeks
Also available as part of
Thrombophilia Screen
2-3
weeks
Also available as part of
Coagulation Screen.
Underfilled, haemolysed,
lipaemic sample will be
rejected.
Presence of anticoagulant
< 4hrs
Male:
0.67 – 139 iu/ml
PROTEIN S
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
Female in
absence of oral
contraception,
HRT or
pregnancy:
060 – 1.13 iu/ml
Fill sample tube to line
indicated on bottle.
PROTHROMBIN TIME
Blood
Citrate
2.7mls
Sample must be
analysed within 4 hours
of the bleed time.
9.3 – 11.8 secs
P a g e | 100
REFERENCE
RANGE
TAT
Negative
Also available as part of
Thrombophilia Screen
2-6
weeks
TEST
SAMPLE
SAMPLE TUBE
VOLUME
PT20210A
Blood
EDTA
4mls
REPTILASE
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
15 – 18 secs
< 4hrs
THROMBIN CLOTTING
TIME
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
14 – 22 secs
< 4hrs
Citrate
4 x 2.7mls
Fill sample tube to line
indicated on bottle
See individual
analytes
THROMOPHILIA
SCREEN
Blood
UNFRACTIONATED
HEPARIN ASSAY
PRECAUTIONS
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
See guidelines for
Thrombophilia Screening
2-3
weeks
EDTA
1 x 4mls
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
Treatment
Blood
VON WILLEBRAND
ACTIVITY
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.7 – 2.0 iu/ml
2-3
weeks
VON WILLEBRAND
ANTIGEN
Blood
Citrate
2.7mls
Fill sample tube to line
indicated on bottle
0.7 – 2.0 iu/ml
2-3
weeks
VON WILLEBRAND
STUDIES
Blood
Citrate
2 x 2.7mls
Fill sample tube to line
indicated on bottle
See individual
analytes
2-3
weeks
0.5 – 1.0 iu/ml
< 4hrs
P a g e | 101
TEST
SAMPLE
SAMPLE TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
By arrangement with
medical staff
1 week
HAEMATO-ONCOLOGY
BONE MARROW
INVESTIGATION
Bone
Marrow
By arrangement
with medical
staff
CD34
Blood
EDTA
4mls
By arrangement with
medical staff
1 week
Venous
Blood
EDTA
4mls
By arrangement with
medical staff
1 week
CELL MARKERS
(PERIPHERAL BLOOD)
For investigation of
haematological
malignancies
NB
Lymphocyte subsets
for investigation of
immunodeficiency
states – refer to
Immunology Lab.
P a g e | 102
TEST
SAMPLE
SAMPLE TUBE
VOLUME
CELL MARKERS
Bone
Marrow
RPMI/Heparin
3 – 5mls
(BONE MARROW)
HEREDITARY
SPHEROCYTOSIS
Blood
EDTA
4mls
Blood
EDTA
4mls
PRECAUTIONS
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
By arrangement with
medical staff
1 week
Negative
MUST be pre-booked by
contacting flow cytometry
staff
1 week
Not detected
By arrangement with
medical staff
1 week
REFERENCE
RANGE
(EMA BINDING STUDY)
PNH SCREEN
See guidelines on PNH
screening
MOLECULAR RED CELL DISORDERS
GAP-PCR
ALPHA
THALASSAEMIA
Blood
BETA THALASSAEMIA
Blood
EDTA
>1ml
PCR direct sequencing
4 weeks
Blood
EDTA
>1ml
PCR direct sequencing
4 weeks
EDTA
>1ml
4 weeks
PCR direct sequencing
ENZYMOPATHIES
(PK AND G6PD)
P a g e | 103
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
ERYTHROCYTOSIS
Blood
EDTA
4mls
PCR direct sequencing
See guidelines in appendix
4.
4 weeks
HAEMOCHROMATOSIS
(HFE)
Blood
EDTA
4mls
PCR
See guidelines in appendix
6.
4 weeks
HAEMOGLOBIN
VARIANTS
Blood
EDTA
>1ml
PCR direct sequencing
4 weeks
JAK2 V617F
Blood
EDTA
>1ml
Fragment Analysis
3 weeks
JAK2 EXON 12
Blood
EDTA
>1ml
High resolution melt
4 weeks
MPL EXON 10
Blood
EDTA
>1ml
High resolution melt
4 weeks
METHAEMOGLOBINAE
MIA
Blood
EDTA
>1ml
PCR direct sequencing
4 weeks
P a g e | 104
MOLECULAR HAEMOSTASIS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
HAEMOPHILIA A
Blood
EDTA
2mls
PCR direct sequencing
F8 gene; inverse PCR
6 weeks
HAEMOPHILIA B
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
TYPE 1 VON
WILLEBRAND
DISEASE
Blood
EDTA
2mls
PCR direct sequencing
6 weeks
TYPE 2 VON
WILLEBRAND
DISEASE
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
TYPE 3 VON
WILLEBRAND
DISEASE
Blood
EDTA
2mls
PCR direct sequencing
6 weeks
INHERITED FACTOR
VII DEFICIENCY
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
TAT
P a g e | 105
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TEST
SAMPLE
SAMPLE
TUBE
VOLUME
PRECAUTIONS
INHERITED FACTOR X
DEFICIENCY
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
INHERITED FACTOR XI
DEFICIENCY
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
DYSFIBRINOGENAEMI
A COMMON
MUTATIONS
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
INHERITED TTP
(ADAMTS13)
Blood
EDTA
2mls
PCR direct sequencing
4 weeks
BERNARD-SOULIER
SYNDROME
Blood
EDTA
2mls
PCR direct sequencing
6 weeks
HHT
Blood
EDTA
2mls
PCR direct sequencing
6 weeks
TAT
P a g e | 106
MOLECULAR HAEMATO-ONCOLOGY
TEST
TRANSLOCATION
ASSAYS
[t(14;18) + t(11;14)]
BCR-ABL
(Monitoring)
BCR-ABL
(Diagnosis)
PCR DIRECT
SEQUENCING
IGVH MUTATION
SCREEN
SAMPLE
Paraffin
embedded
tissue,
fresh
tissue,
BM/PB
Blood –
EDTA
Blood –
EDTA or
Bone
marrow
Blood –
EDTA/
Bone
marrow
SAMPLE
TUBE
VOLUME
PRECAUTIONS
5 x 10um
paraffin
processed
sections of
tissue, 5 ml
BM/PB
PCR
10- 20 ml PB
10- 20 ml PB
or 1-3ml BM
1-3 BM and
5ml-PB
It is important that these
samples reach the
laboratory within 24
hours of being taken as
any samples received
after this time will not be
processed as RNA
quality cannot be
guaranteed.
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
2 weeks
See guidelines in
appendix 11.
2 weeks
See guidelines in
appendix 11.
2 weeks
2 weeks
P a g e | 107
SAMPLE
TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TEST
SAMPLE
TAT
IMMUNOGLOBULIN
GENE
REARRANGEMENTS
(PCR)
Paraffin
embedded
tissue,
fresh
tissue,
BM/PB
5 x 10um
paraffin
processed
sections of
tissue, 5 ml
BM/PB
2 weeks
T-CELL RECEPTOR
GENE
REARRANGEMENTS
(PCR)
Paraffin
embedded
tissue,
fresh
tissue,
BM/PB
5 x 10um
paraffin
processed
sections of
tissue, 5 ml
BM/PB
2 weeks
P a g e | 108
Referral Laboratories
Blood Bank
Test: Antibody Investigation
1. Northern Ireland Blood Transfusion Service (NIBTS)
Belfast City Hospital Complex
Lisburn Road
Belfast
BT9 7TS
Tel: 028 9032 1414
Haematology
Test: Plasma Viscosity
2. The Ulster Hospital
Upper Newtownards Road
Dundonald
Belfast
Co Down
BT16 1RH
Tel: 028 9048 4511
P a g e | 109
APPENDICES
The following provide additional information or useful guidance referred from the
individual test information or the laboratory reports.
Appendix 1: Sickling disorders - Sickle solubility testing
Appendix 2: WBC in Africans and Caribbeans of African Lineage
Appendix 3: Reporting of Blood Films by Medical Staff
Appendix 4: Guidelines for Red Cell Volume Investigation
Appendix 5: Bone Marrow Aspirate and Trephine Biopsy
Appendix 6: Guidelines for HFE Gene Screening
Appendix 7: Methylene blue fresh frozen plasma or cryoprecipitate
Appendix 8: Provision of HLA selected platelet components for alloimmunised
patients
Appendix 9: Clinical Significance of Red Cell Antibodies
Appendix 10: B-cell Chronic Lymphocytic Leukaemia: Molecular Analysis
Appendix 11: Updated Guidance: BCR/ABL Detection, Quantitative Monitoring
& Mutation Analysis in CML
Figure 1:
Flow chart for monitoring CML patients on TKI therapy
Appendix 12: Directions for Obtaining a Specimen for Post Vasectomy
Investigation
P a g e | 110
Appendix 1:
Department of Haematology, Belfast City Hospital
Sickling disorders - Sickle solubility testing
Sickle solubility testing is available at RVH and BCH labs. This is a rapid screening
test. A positive result indicates the presence of HbS but does not distinguish
between sickle cell disease (Hb SS), sickle cell trait (Hb AS), and various compound
heterozygous states. In the acute setting, before confirmatory results are available,
one should manage the patient as sickle cell disease.
(a) Positive sickle solubility test and normal blood film: Assume sickle cell trait.
(b) Positive sickle solubility test and any sickle cells or target cells on blood film:
Assume sickle cell disease, irrespective of Hb (for example patients with HbSC
disease may have a normal Hb but still have a clinically significant acute sickling
crisis).
N.B. Sickle solubility tests are often negative in infants with sickle cell disease (due
to the protective effect of HbF).
False positive sickle solubility results:




Severe leucocytosis
Hyperproteinaemia, eg myeloma
Hyperlipidaemia
Unstable haemoglobins – especially after splenectomy
False negative sickle solubility results:



Low Hb
Infants <6 months
Post-transfusion
RJG Cuthbert,
Last updated August 2014
Adapted from Practical Haematology, 10th Edition, Churchill Livingstone, 2006
P a g e | 111
Appendix 2:
Department of Haematology, Belfast City Hospital
WBC in Africans and Caribbeans of African Lineage
95 percentile ranges (x109/L)
Male
Female
Origin
WBC
Neut
WBC
Neut
African
2.8-7.2
0.9-4.2
3.0-7.4
1.3-3.7
Carribbean
3.1-9.4
1.2-5.6
3.2-10.6
1.3-7.1
From Barbara Bain, Blood Cells: A Practical Approach, 2006
P a g e | 112
Appendix 3:
Department of Haematology, Belfast City Hospital
Reporting of Blood Films by Medical Staff
Introduction: Examination of a blood film is an essential procedure in the clinical
assessment, investigation, and interpretation of abnormal FBC results. Initial
screening is undertaken by qualified Biomedical Scientist staff, who will refer
samples to the lab registrar:
(i)
(ii)
In new patients where there is a difficulty in interpretation
In cases of suspected primary haematological disorders.
Specimen Preparation: Blood films are spread from fresh EDTA samples in the
laboratory. They should be prepared within two hours (but not exceeding 12 hours)
of blood collection. Well-spread, well-stained films are required to ensure reliable
information can be acquired.
Methods: A systematic approach to blood film examination is essential to correct
interpretation.




FBC samples are selected for blood film examination according to numerical
criteria, analyser flags, and clinical flags
The blood film is referred by Biomedical Scientist staff along with the FBC
request form &/or the morphology referral form
Referrals are placed in a tray basket on the registrar’s bench in the lab
The lab registrar should liaise regularly – at least twice daily - with BMS staff, to
look out for samples requiring urgent action
Results: Interpretative comments are entered into LabCentre. Clinical interpretation
is dependent on the patient’s history, FBC results, other investigations, and
comprehensive knowledge and experience of clinical haematological practice.
Therefore, inexperienced registrars must discuss all cases with the duty lab
consultant.
References:


Bain, BJ. Morphology of blood cells. Ch 3. pp 61-174. In: Blood Cells: A
Practical Guide. Blackwell Publishing Ltd, 4th Edition, 2006.
Bain, BJ. Blood film morphology in health and disease. Ch5, pp 79-113. In:
Practical Haematology. Churchill Livingston Elsevier Ltd, 10th Edition, 2006.
Department of Haematology
Belfast City Hospital
Updated April 2012
P a g e | 113
Appendix 4:
Department of Haematology, Belfast City Hospital
Guidelines for Red Cell Volume Investigation
Red cell volume investigation involves iv administration of radiolabelled autologous
red cells. It is undertaken to diagnose or exclude absolute erythrocytosis in patients
who are negative for JAK-2 V617F mutation.
Indications:
1. Packed cell volume (haematocrit) elevated for >2 months:
Male
PCV >0.52
Female PCV >0.48
Minimal or no venous occlusion when taking the blood sample
2. Erythrocytosis may be masked by Fe deficiency:
Typical FBC:
Hb usually upper end of normal range
PCV usually upper end of normal range
Low MCV
Raised RCC
Fe replacement should be undertaken only with extreme caution
PCV may rise rapidly and precipitate thrombosis
Monitor Hb and PCV weekly
Not indicated:
1. PCV normal (unless 2 above applies)
2. PCV grossly elevated:
Male:
PCV >0.60
Female
PCV >0.56
These patients have absolute erythrocytosis
P a g e | 114
Requesting the Investigation:
Requests should be submitted to:
Nuclear Medicine Department
Level 1 Imaging Centre
Royal Victoria Hospital
Use the appropriate radiology request form, and please submit giving full clinical
details.
RJG Cuthbert
Updated February 2014
This document is adapted from: Guidelines for the diagnosis, investigation and
management of polycythaemia/erythrocytosis Br J Haematol, 2005;130:174-9.
P a g e | 115
Appendix 5:
Department of Haematology, Belfast City Hospital
Bone Marrow Aspirate and Trephine Biopsy
Introduction: As part of the investigation of patients with a known or suspected
haematological disorder, it is often necessary to perform a bone marrow aspirate and
trephine biopsy procedure. The duty laboratory registrar undertakes the initial
assessment and the results are scrutinised by the consultant haematologist before
final authorisation.
Specimen Collection and Preparation: Bone marrow aspirate slides are made at
the patient’s bedside using pink frosted glass slides and labeled with the patient’s
first name & surname, and the date of the procedure. Aspirate samples for flow
cytometry and cytogenetics are placed in universal containers containing RPMI and
heparin. Aspirate samples for molecular studies are placed in EDTA tubes. The
trephine biopsy sample, if taken, is placed in a 5mL plain plastic tube containing 10%
formalin labelled appropriately. The samples are delivered to the bone marrow/flow
cytometry laboratory for processing.
Principles of Methods: Various stains can be used to visualise blood cells and their
precursors. These include Wright’s stain and Perl’s stain for haemosiderin
performed on the aspirate smears, and H&E and Giemsa stains performed on the
trephine sections. The stained slides are assessed, the various cell populations
enumerated and then reported by suitably trained medical staff. The results help in
the diagnosis and management of patients. Immunonhistochemistry has an
important role in trephine diagnosis, but is expensive.
Judicious use of
immunonhistochemistry may be of diagnostic benefit to patients but must be used
systematically.
Clinical Interpretation: Examination of Wright’s stained bone marrow aspirate
slides, assessment of iron stores, examination of trephine sections and cytochemical
stains is a complex procedure that requires specialist training and experience. It can
affect a patient’s diagnosis and treatment and as such should only be performed by
appropriately qualified staff.
References:
Lee S-H et al. ICSH guidelines for the standardization of bone marrow specimens
and reports. Int J Lab Haematol, 2008; 30: 349-64
P a g e | 116
Appendix 6:
Department of Haematology, Belfast City Hospital
Guidelines for HFE Gene Screening
Individuals with symptoms or signs for which haemochromatosis might be
considered in the differential diagnosis are candidates for investigation. Early
symptoms of haemochromatosis may be non-specific. Thus genetic testing is not
appropriate without first assessing body iron status. Widely accepted criteria for
excessive body iron are as follows:

Serum ferritin
>300 μg/L in males
>200 μg/L in females

Fasting transferrin saturation >55%
Transferrin saturation should be assessed in the fasting state since serum iron is a
physiologically labile analyte. Since ferritin is an acute phase reactant, the results of
iron profile investigations must be interpreted within the context of the individual’s
clinical status.
Individuals with a positive family history of clinically apparent haemochromatosis
have a higher risk of developing overt disease and, therefore, are candidates for
investigation.
Criteria for HFE mutation screening:


Patients with evidence of iron overload and clinical features suspicious of
haemochromatosis.
First degree relatives of an index case who is homozygous for C282Y or
compound heterozygous for C282Y/H63D.
Sample to send: Please send a routine EDTA sample (FBC tube) and haematology
request form with full clinical details to:
Haematology Laboratory,
Belfast City Hospital,
Lisburn Road,
Belfast, BT9 7AB
Lab. tel. no. 02890-329241 Ext. 3097
P a g e | 117
What to do with the most commonly encountered abnormal results:






C282Y heterozygote with normal iron studies - no action required.
H63D heterozygote with normal iron studies - no action required.
C282Y homozygote with normal iron studies - check iron status every 1-2
years.
C282Y/H63D compound heterozygote with normal iron studies - check iron
status every 1-2 years.
C282Y homozygote with raised iron stores - refer according to your usual
referral patterns
C282Y/H63D compound heterozygote with raised iron stores - refer according
to your usual referral patterns
Dr RJG Cuthbert,
Updated July 2015
P a g e | 118
Appendix 7:
Department of Haematology, Belfast City Hospital
Methylene Blue Fresh Frozen Plasma or Cryoprecipitate
Anyone born from the 1st January 1996
Cytomegalovirus (CMV) negative red cells



CMV negative red cells and platelet components should be provided for intra
uterine transfusions and for babies up to the age of 20 weeks
CMV negative blood components should be provided where possible for
women, during pregnancy regardless of their CMV status (this applies during
pregnancy but not labour or delivery). For emergency transfusions in this
group , where CMV negative products are not readily available ,
leucodepleted components are recommended
Granulocytes components should be CMV negative for CMV seronegative
patients
Irradiated red blood cells











Red cells for intrauterine transfusions
Neonatal exchange transfusions
Severe T lymphocyte immunodeficiency syndromes
Top up transfusions if the baby has received intrauterine transfusions
Bone marrow, stem cell transplant patients, and donors of bone marrow /
stem cell
Hodgkin’s lymphoma
Aplastic anaemia receiving ATG (and/or alemtuzumab)
Patients on purine analogue (type of chemotherapy) drugs (fludarabine,
cladrabine and deoxycoformycin)
Patient’s on bendamustine, clofarabine, alemtuzumab
HLA (human leucocyte antigen) selected units
Transfusion from first or second degree relative
HLA matched red cells are indicated for:


Patients awaiting a renal transplant
Those who have been transplanted
immunosuppressed.
in
the
past
and
are
non-
This is best practice however if in an emergency situation it is down to a clinical
decision if HLA is not available.
Patient groups for whom HLA matched platelets are indicated:


Platelet dysfunction in chronic renal failure/uraemia
Patients who have already demonstrated HLA antibodies and who may show
platelet refractoriness
P a g e | 119
Appendix 8:
Department of Haematology, Belfast City Hospital
Provision of HLA selected platelet components for alloimmunised
patients
Platelet refractoriness is defined as two consecutive failures of response to platelet
transfusions, i.e. failure to achieve 24 hour post transfusion platelet count > 20 x
109/l.
The majority of cases of platelet refractoriness are due to non-immune causes.
These have been identified in studies by Bishop et al from the Melbourne Blood
Centre and published in Transfusion as DIC consumption, anti-microbial therapy
especially amphotericin B, pyrexia, hypersplenism, post BMT (immune
dysregulation). Non-immune causes should be treated to optimise response to
platelet substitution therapy.
Immune causes which account for no more than 25% of cases are anti-ABO
antibodies, anti-platelet specific antibodies and anti-HLA antibodies.
The first line of treatment should be ABO identical platelet components with high
dose (NIBTS platelet components have the platelet yield on the label). Our quality
monitoring data demonstrate average yield for single donor platelets (apheresis) of
280 x 109/l but with reduced results for pools of buffy coat derived platelets of 238 x
109/l.
Where anti-platelet specific antibodies are identified, NIBTS will endeavour to
provide HPA specific antigen negative platelet components.
Where anti-HLA antibodies are identified, NIBTS will search for matching donors but
we often have to make use of selected mismatching because of lack of HLA identical
or HLA homozygous haplotype donors.
It is imperative that NIBTS medical team receive follow up information on increment
data and clinical response when HLA selected components are transfused. This will
enable us to target donors and provide optimum support.
Therefore the following platelet support strategy will be adopted:

Transfuse ABO identical platelet components (single donor platelets
preferred).

Select high yield dose single donor platelets.

Provide HPA specific platelet components if appropriate.
P a g e | 120


Provide HLA selected platelet components if appropriate. This would normally
require confirmation of HLA antibodies and 3 month clinical assessment follow
up.
Return increment clinical response data to NIBTS medical team periodically.
Prepared by: Dr Morris and Dr Maguire Date: 10 September 2014
MP14/02
P a g e | 121
Appendix 9:
Clinical Significance of Red Cell Antibodies




Clinically significant antibodies are those that are capable of causing patient
morbidity due to the accelerated destruction of a significant proportion of
transfused red cells.
Anti-A, anti-B and anti-A,B must always be regarded as being of clinical
significance.
With few exceptions, red cell antibodies which are likely to be of clinical
significance are only those which are reactive in the indirect antiglobulin test
(IAT), performed strictly at 37oC.
Recommendations for the selection of red cells for transfusion to patients with
alloantibodies are given in table 1 below.
Table 1 - Likely clinical significance of red cell alloantibodies, and
recommendations for the selection of blood for patients with their presence
System
Specificity
Likely clinical
significance
in transfusion
Recommendation for selection of red
cells for transfusion *
ABO
Anti-A1
No
IAT crossmatch compatible at 37oC
Rh
Anti-D, -C, -c, -E, -e
Yes
Antigen negative
Rh
Anti-Cw
No
IAT crossmatch compatible **
Kell
Anti-K, -k
Yes
Antigen negative
Kell
Anti-Kpa
No
IAT crossmatch compatible **
Kidd
Anti-Jka, -Jkb
Yes
Antigen negative
MNS
Anti-M (active 37oC)
Yes
Antigen negative
MNS
Anti-M (not active 37oC)
No
IAT crossmatch compatible at 37oC
MNS
Anti-N
No
IAT crossmatch compatible at 37oC
MNS
Anti-S, -s, -U
Yes
Antigen negative
P a g e | 122
Duffy
Anti-Fya, -Fyb
Yes
Antigen negative
P
Anti-P1
No
IAT crossmatch compatible at 37oC
Lewis
Anti-Lea, -Leb, -Lea+b
No
IAT crossmatch compatible at 37oC
Lu
Anti-Lua
No
IAT crossmatch compatible at 37oC
Diego
Anti-Wra (anti-Di3)
Yes
IAT crossmatch compatible **
H
Anti-HI (in A1 and A1B patients)
No
IAT crossmatch compatible at 37oC
All
Others active by IAT at 37oC
Yes
Seek advice from Blood Centre
* Where antigen negative red cells are recommended these should also be
compatible in an IAT crossmatch.
** These recommendations apply when the antibody is present as a sole specificity.
If present in combination, antigen negative blood may be provided by the blood
centre, to prevent wastage of phenotyped units.
The above guidance is also suitable for patients undergoing hypothermia during
surgery (Mollison, 2005b).
From BCSH Guidelines for Pre-Transfusion compatibility procedures in Blood
Transfusion Laboratory, 2012.
P a g e | 123
Appendix 10:
Department of Haematology, Belfast City Hospital
B-cell Chronic Lymphocytic Leukaemia:
Molecular Analysis
The cytogenetics laboratory at Belfast City Hospital is now offering prognostic testing
by FISH for new patients with B-CLL. The analysis using a Vysis kit will include:




del 17p13 (p53 deletion)
del 11q22-23 (ATM)
del 13q14.3
trisomy 12
The BCH haemato-oncology laboratory will undertake investigation of somatic
hypermutation status in IG genes.
The criteria for testing will remain the same as those used to date for referral to
HMDS at Leeds:




Pre-treatment sample
WBC>50x109/L, or
Patient requires treatment
Refractory/relapse patient at discretion of referring haematologist
To help in the differential diagnosis of B-CLL and mantle cell lymphoma
t(11;14)(q13;q32) will be investigated by FISH in cases with atypical morphology
and/ or an atypical immunophenotype. (CD200 expression is under development,
and we shall send a further circular about this very soon.)
Peripheral blood samples drawn into EDTA are preferred to bone marrow. However,
for small lymphocytic lymphoma without a leukaemic phase it will be necessary to
process bone marrow and/ or fresh lymph node specimens.
P a g e | 124
Samples should be sent as usual for flow cytometry to the haemato-oncology
laboratory at Belfast City Hospital with full clinical details. Samples will be CD19enriched and sent to the cytogenetics laboratory and the haematology molecular
section based on the above criteria.
Please do not send samples directly to the cytogenetics laboratory as this is
generating unnecessary duplication of work.
Mervyn Humphreys, Gary Beattie, Mark Catherwood, Robert Cuthbert
Updated August 2015
P a g e | 125
Appendix 11:
Department of Haematology, Belfast City Hospital
Updated Guidance: BCR/ABL Detection, Quantitative Monitoring
& Mutation Analysis in CML
Diagnosis: The diagnosis of CML is usually suspected by an elevated WBC with left
shift, basophilia and splenomegaly. Whilst detection of peripheral blood BCR/ABL
by RT-PCR confirms the diagnosis, bone marrow aspiration is still recommended to
establish the baseline blast count. This allows differentiation of chronic, accelerated,
and blast phases of the disease. BM cytogenetics is important in identifying
additional chromosome abnormalities at diagnosis.
A baseline RT-PCR result is essential in planning subsequent therapeutic
monitoring. Without it, uncommon rearrangements may not be identified after
initiation of treatment, and so molecular monitoring could be compromised.
Monitoring treatment: During early treatment frequent clinical evaluation + FBC is
essential to establish clinical/haematological response, and identify potential toxicity.
Patients who fail to achieve a complete haematological response at 3 months should
be considered for second-line therapy. A PB sample should be sent at this stage for
BCR/ABL mutation analysis.
Patients achieving a complete haematological response should have a PB sample
sent for RT-qPCR at 3 months and BM aspirate for cytogenetics at 6 months.
Subsequent strategy is dependent on the results of the 6 month assessment (see
Figure 1):
(a) Complete cytogenetic response (CCyR): Further routine BM cytogenetics is
not required. PB samples should be sent for RT-qPCR at 3 month intervals until a
major or complete molecular response is achieved. Subsequently, PB samples
should be sent for RT-qPCR at 6 month or longer intervals.
(b) Major cytogenetic response: Treatment should be continued at the standard
dose and BM cytogenetics repeated at 3-6 month intervals. A significant minority of
patients will achieve a CCyR, and can then join group (a). Achievement of CCyR
remains the gold standard for defining an adequate response because it is
associated with prolonged survival.
P a g e | 126
(c) Failure to achieve CCyR: Patients who fail to achieve CCyR at 18 months
should be considered for second-line therapy. A PB sample should be sent at this
stage for BCR/ABL mutation analysis.
(d) Major or complete molecular response: At 18 months patients who achieve >3
log reduction in BCR/ABL copies, or have undetectable BCR/ABL using a highly
sensitive RT-qPCR assay, have a modestly improved EFS compared with those in
CCyR without a MMR.
(e) Failure to achieve MMR: Patients in CCyR who fail to achieve >3 log reduction
should continue on treatment. They should be considered for follow up BM
cytogenetics at 3-6 month intervals. If CCyR is lost the patient should be considered
for second-line treatment. A PB sample should be sent at this stage for BCR/ABL
mutation analysis.
(f) Loss of previously achieved MMR/CMR: Patients who subsequently lose a
MMR or CMR (>1 log increase, ie – 0.2-2% BCR/ABL transcripts) should be carefully
assessed for treatment compliance, and have repeat BM cytogenetics. If CCyR is
maintained treatment should not be changed. However, follow up BM cytogenetics
at 3-6 month intervals is indicated. If CCyR is lost the patient should be considered
for second-line treatment. A PB sample should be sent at this stage for BCR/ABL
mutation analysis.
Kinase Domain Mutation Analysis: Mutations within the BCR-ABL1 kinase domain
(KD) may cause acquired resistance to imatinib. Kinase domain mutation analysis is
now available in the regional molecular haematology laboratory. Criteria for mutation
analysis include primary TKI-refractory disease, failure to achieve a major molecular
response within 18 months of commencing TKI therapy, loss of MMR or presentation
with advanced disease. Mutational analysis is not performed at diagnosis.
Sending Samples to the Lab.
PB samples (minimum 12mL) in EDTA must arrive in the laboratory within 24 hours
of sampling. Full clinical details are essential.
Send samples to: Department of Haematology,
C Floor, Belfast City Hospital,
Lisburn Road, Belfast, BT9 7AB
Contact details:
Scientific advice: Dr Mark Catherwood – 028-950-40914
Clinical interpretation: Dr RJG Cuthbert – 028-950-47989
P a g e | 127
Abbreviations/Definitions:
Complete haematological response:
Plt<450
No immature granulocytes
WBC <10.0 No residual basophilia
No splenomegaly
RT-qPCR – Quantitative PCR technique to determine of the number of BCR/ABL
transcripts
CCyR – Complete cytogenetic response
MCyR - Major cytogenetic response - <35% Ph’ chromosomes in bone marrow
CMR – Complete molecular response
MMR – Major molecular response - >3 log reduction in number of BCR/ABL
transcripts, ie <0.1%
RJG Cuthbert & M Catherwood October 2012 (reviewed July 2015)
P a g e | 128
Figure 1:
Flow chart for monitoring CML patients on TKI therapy
1. Blood film
BM cytogenetics
RT-PCR
2. Haematological response
Yes
No
3. RT-qPCR at 3/12
15. Change Rx
Mutation analysis
3. BM cytogenetics at 6/12
4. CCyR
No
Yes
No
5. RT-qPCR q3/12
12. Continue Rx
BM cytogenetics at 12/12
& if necessary at 18/12
13. CCyR
Yes
6. MMR or CMR
Yes
No
11. Continue Rx
Repeat BM cytogenetics q3-6/12
14. Continue Rx
Go to 5.
7. RT-qPCR q6/12
9. Loss of response
8. Continue
Rx
10. Continue Rx
Repeat BM cytogenetics
RJG Cuthbert & M Catherwood October 2012 (reviewed July 2015)
P a g e | 129
Appendix 12:
Department of Haematology, Belfast City Hospital
Andrology: “Directions for obtaining a specimen for post
vasectomy investigation”

The patient should have abstained from sexual activity (intercourse or
masturbation) for a minimum of 2, but no longer than 7, days before submitting
a specimen.

A special sterile wide-mouth plastic container is provided.

To avoid bacterial contamination the patient should wash his hands and penis
3 times in warm soapy water, rinsing well before drying with soft tissues.

A complete specimen of semen obtained by masturbation should be allowed to
run into the container and the cap firmly replaced.

After producing the specimen, the patient should complete and check the
relevant section of the request form and sign to confirm that the information is
correct. The patient should complete and check the specimen container label,
including the time and date the specimen was obtained.

If the patient has difficulty producing a sample he should let the laboratory know
and arrange another appointment.

Immediately after collection the specimen should be delivered to the
Haematology Laboratory. Please note: there are no facilities available for:
 Obtaining a specimen or
 The production of a specimen at the laboratory

The specimen should be at the laboratory no later than 1 hour after
collection.
During this time the specimen should be kept at body temperature e.g. by placing
the container in an inside pocket of the clothes being worn. This is because
cooling may affect the outcome of analysis.
If there is a delivery time of more than 1 hour please notify laboratory staff on
arrival.
P a g e | 130
REGIONAL HISTOCOMPATIBILITY & IMMUNOGENETICS
General Information:
The Belfast Health & Social Care Trust (BHSCT) Histocompatibility &
Immunogenetics (H&I) Laboratory is a regional speciality laboratory which provides a
range of highly complex testing. The laboratory is located on the Belfast City Hospital
(BCH) site within the Northern Ireland Blood Transfusion (NIBTS) building.
The full postal address for the H&I Laboratory is:
Regional Histocompatibility & Immunogenetics Laboratory
First Floor, Northern Ireland Blood Transfusion Building
Belfast City Hospital,
Lisburn Road
Belfast
BT9 7TS
Services Offered:
The laboratory provides services which may be considered under two main areas:
Renal Services

Non Renal Services
The laboratory plays a critical role in renal transplantation (living and deceased
kidney donation) by undertaking tests to ensure optimum human leukocyte antigen
(HLA) matching between donors and patients and antibody screening and cross
matching to minimise the likelihood of graft rejection and failure.
Non renal services includes haematopoietic stem cell transplantation, transfusion
related services and genetic association testing. The laboratory plays a vital role in
related Bone Marrow transplantation by undertaking tests to ensure optimum
matching between donors and patients. A service level agreement exists with the
NIBTS, in which the laboratory provides a HLA typing service for volunteers
associated with the British Bone Marrow donor registry.
The main genetic association test which is currently undertaken by the Laboratory is
HLA-B27 testing, principally for Ankylosing Spondylitis.
P a g e | 131
Opening Hours:
Normal working hours are 9.00 am to 5.00 pm Monday to Friday, excluding bank
holidays.
The H&I laboratory provides a 24 hour, 365 days per year out-of-hours on call
service for renal transplantation. Enquiries to the duty on-call scientist are made via
the Belfast Trust switchboard.
Contact Details:
H&I staff can be contacted as follows:



Office: 028 950 43240 (43240)
Analogue contingency Line: 208 906 37771
Fax No: 028 906 37741
The main contacts are:
Consultant/Head of Laboratory:- Dr. David Savage BCH Ext 44045
Tel: 028 950 44045
Email:david.savage@belfasttrust.hscni.net
H&I Service Manager:-Miss Elaine Boyle BCH Ext 44529
DDI: 028 950 44529
Email:elaine.boyle@belfasttrust.hscni.net
Senior Clinical Scientist:- Miss Bernadette Magee BCH Ext 43702
DDI: 028 950 43702
Email:bernie.magee@belfasttrust.hscni.net
Senior Clinical Scientist:- Dr. Brian McIlhatton BCH Ext 42893
Email:brian.mcilhatton@belfasttrust.hscni.net
Serology Section Lead:- Mr. Miceal Cole - BCH Ext 42894
Email:miceal.cole@belfasttrust.hscni.net
Serology Section Lead:- Mr. Ashley Meenagh - BCH Ext 42894
Email:ashley.meenagh@belfasttrust.hscni.net
Molecular Services Lead:- Mr. Michael McDermott - BCH Ext 42893
Email:michael.mcdermott@belfasttrust.hscni.net
Clinical Advice:
When advice on the clinical relevance of laboratory results is required out of hours
please contact the on-call duty clinical scientist or consultant/head of laboratory via
the BHSCT switchboard.
P a g e | 132
Request Form & Sample Labelling:
Request forms should contain the following essential information:
1.
2.
3.
4.
5.
6.
7.
8.
Patients full name (first name and surname)
Date of birth
Gender
Health & Care Number and / or Hospital Number
Date sample taken
Test request
Include name and contact details of requesting clinician
Signed by clinician, nurse or phlebotomist who has taken the sample
Each sample must be appropriately labelled, accompanied by a request
form and contain the following essential information:
1. Patients full name (first name and surname)
2. Date of birth
3. Health & Care Number and / or Hospital Number
4. Date sample taken
5. Signed by clinician, nurse or phlebotomist who has taken the sample.
Note: to confirm that the patient’s details have been correctly recorded on the
request form and sample tube, all renal samples and forms must be double
signed by staff involved in sampling. When a patient is an adult and judged
competent to do so, he or she may be the second signatory.
Users should be aware that failure to meet these requirements may result in
sample rejection
Requirements for Transport of Samples
Users should refer to the Trust policy document titled ‘Transport of specimens by
road’ policy C-18/LI 800 026. This is located in the Belfast Trust Laboratories (BTL)
User Manuals which is accessible on the home page of the intranet and going to the
drop down option of ‘View the Labs User Manual’ in the ‘I want to…’ section.
Or on the internet at:
http://www.belfasttrust.hscni.net and in the Services Tab select Adult Services,
followed by Laboratory and Mortuary Services & moving the cursor over ‘Belfast
Trust Laboratories BTL User Manual’ (purple section) under the heading
Laboratories.
Communication of Results
The H&I laboratory does not directly report results to patients/parents or carers. All
results are reported to the requesting clinician/GP.
P a g e | 133
User Satisfaction
A management review (AMR) is held within the H&I laboratory annually. This serves
to identify any changes required to meet the needs and requirements of users, and
any action needed to ensure the continuation of the service. An approved version of
the AMR report will be made available to users of the service upon request to the
laboratory manager.
A user satisfaction survey is conducted regularly by the H&I laboratory and forms
part of the AMR. Current users of the H&I laboratory will be invited to participate in
this survey and responses are discussed, acted upon where appropriate and
included in the AMR.
P a g e | 134
TEST
SAMPLE SAMPLE TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Within
90%
Routine HLA
Molecular Typing for
Renal Recipient &
Living Donor
Blood
Purple Top EDTA
9 ml
N/A
N/A
N/A
Up to 21
days
Routine HLA
Molecular Typing for
Haematopoietic Stem
Cell Transplantation
(HSCT) Recipient and
Potential Donors
Blood
Purple Top EDTA
4 ml
All HLA-typing of families N/A
within N. Ireland must be
pre-booked and organised
through the Bone Marrow
Transplant
Co-ordinator
(via BCH switchboard –
Bleep No. 1675).
N/A
Up to 21
days
N.B. For potential
recipients living outside N.
Ireland with family
members living in N.
Ireland requiring HLA
typing, the request must be
accompanied by a
statement of authorisation
from the patient’s Health
Authority that they will pay
for testing.
P a g e | 135
TEST
All Living Donor
Crossmatching
1. Recipient
(Auto
Crossmatch)
SAMPLE
Blood
and
Blood
SAMPLE TUBE
Light Blue
Sodium Citrate
60ml
Clotted Red Top
gel tube
9ml
Blood
2. Donor
Routine HLA
Antibody Screening
VOLUME
PRECAUTIONS
To be organised via Living
Donor Transplant Coordinators
BCH#48293/49437
N/A
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
Ideally taken
from peripheral
vein. Sent to
the laboratory
first thing on day
of crossmatch.
60ml
Light Blue
Sodium Citrate
Blood
REFERENCE
RANGE
Clotted Red Top
gel tube
N/A
9ml
Routine HLA antibody
screening to be performed
every 3 months for all
patients active and
suspended on the waiting
list, and from all patients
identified as suitable for
transplantation.
N/A
Ideally the
sample should
be received in
the lab within 24
hrs.
TAT
Within 90%
Within 2
working
days if
donor and
recipient are
HLA typed.
An interim
report will be
provided if
required
when HLA
types are
not
available.
Up to 42
days.
P a g e | 136
TEST
SAMPLE SAMPLE TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Within
90%
URGENT HLA
Antibody Screening &
Identification
Blood
Clotted Red Top
gel tube
9ml
To be organised through
the H&I laboratory.
N/A
N/A
5 working
days.
HLA Antibody
Screening
Blood
Clotted Red Top
gel tube
9ml
Samples must be taken on
2 occasions after a blood
transfusion, i.e. ideally 1428 days post-transfusion.
This protocol applies both
to all patients on the
waiting list and to those
identified as suitable for
transplantation.
N/A
N/A
Up to 42
days.
HLA Autoantibody
Crossmatching
Blood
Light Blue
Sodium Citrate
40ml
These samples are
organised by the renal
recipient transplant coordinator on instruction
from the laboratory staff.
N/A
N/A
Within 2
working
days.
HLA Typing for Local
Deceased Donors
Blood
Light Blue
Sodium Citrate
20ml DBD
Must be arranged through
the Specialist Nurses for
Organ Donation (SNOD).
N/A
N/A
Within 6
hours
For Renal Recipients Post-Transfusion
Purple Top EDTA
Clotted Red Top
gel tube
100ml
DCD
20ml
9ml
P a g e | 137
TEST
SAMPLE SAMPLE TUBE
Deceased Donor
Crossmatching
Spleen/
HLA Matched Blood
for Renal Recipients
Blood
HLA Matched
Platelets
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
N/A
N/A
N/A
N/A
N/A
Within 6
hours
Light Blue
Sodium Citrate
9ml
N/A
N/A
Up to 3
working
days.
Purple Top EDTA
4ml
Clotted Red Top
gel tube
9ml
This service is provided
Monday - Friday in
conjunction with NIBTS.
The patient’s consultant
must contact an NIBTS
consultant to discuss the
patient’s requirements
(028 90321414 ext. 4678).
NIBTS liaise with the H&I
lab staff.
Light Blue
Sodium Citrate
9ml
This service is provided
Monday - Friday in
conjunction with NIBTS.
The patient’s consultant
must contact an NIBTS
consultant to discuss the
patient’s requirements
(028 90321414 ext. 4678).
NIBTS liaise with the H&I
lab staff.
N/A
N/A
Up to 3
working
days.
Lymph
Node
Blood
Within
90%
4ml
Purple Top EDTA
Clotted Red Top
gel tube
9ml
P a g e | 138
TEST
SAMPLE SAMPLE TUBE
VOLUME
PRECAUTIONS
REFERENCE
RANGE
KEY FACTORS
AFFECTING
PERFORMANCE /
INTERPRETATION
TAT
Within
90%
HLA Typing for Bone
Marrow & Platelet
Registers
Blood
Purple Top EDTA
6ml
Samples are taken from
potential donors at
N.I.B.T.S. blood donation
sessions, and should
arrive in the laboratory
within 24 hours.
N/A
N/A
Up to 21
days
HLA-B27 Testing
Blood
Purple Top EDTA
4ml
N/A
N/A
N/A
Up to 15
days
Any H&I requests not mentioned above must be discussed with the Head of Laboratory/Discipline Specific Manager
before sending the blood sample.
P a g e | 139
Regional Immunology Service
Introduction
The Regional Immunology Service is based at the Immunology Day Centre and at
the Kelvin Laboratory site, Royal Hospitals, Belfast Health & Social Care Trust.
The laboratory is the only CPA accredited provider of Immunology services in
Northern Ireland.
Our full postal address is:
Clinical Service:
Laboratory Service:
Regional Immunology Service
Regional Immunology Service
Immunology Day Centre,
Kelvin Laboratories,
Royal Hospitals
Royal Hospitals
Belfast Health & Social Care Trust
Belfast Health & Social Care Trust
Grosvenor Road
Grosvenor Road
Belfast
Belfast
BT12 6BA
BT12 6BA
Clinical Service
The clinical Immunology service receives referral in the areas of allergy, immune
deficiency and autoimmune disease.
Adult clinics are held on Level 6 Outpatients Department, Royal Victoria Hospital.
Paediatric outpatient clinics are carried out in the Royal Belfast Hospital for Sick
Children (RBHSC). In addition a joint adult respiratory clinic (lung defence clinic) is
held monthly with Dr D Downey at the Belfast City Hospital. A multispecialty clinic
with paediatric respiratory, infectious disease and immunology, is held monthly at the
RBHSC for assessment and management of complex paediatric cases.
The clinical service provided at the Immunology Day Centre includes infusion clinics
for intravenous immunoglobulin (IVIG) and biological drugs and an IVIG home
therapy service. Allergy challenge testing and allergen desensitisation is also
undertaken.
Clinical referrals are welcomed.
Website http://allergyandimmunologyni.org/
P a g e | 140
Medical staff:
Dr JDM Edgar (Consultant Immunologist)
02890 636649 / 02890 632663
Dr Lisa Devlin (Consultant Immunologist)
02890 631803 / 02890 633853
Dr Hiba Shendi (Consultant Immunologist)
02890 630002
Dr Cathal Steele (Specialist Registrar)
02890 631804
Secretaries
02890 633620am
02890 630007pm
02890 630003
Immunology Day Centre nursing staff:
Denise Florence
02890 630001
Deborah Kealey
02890 630001
Alison Donnelly (Allergy)
02890 636456
Nurses reception
02890 635385
Fax number
02890 635482
Laboratory Service
The laboratory is open from 9-00 am to 5-15 pm Monday to Thursday and 9-00 am
to 5-00 pm on Friday.
Any out of hours requests should be directed to the Royal Hospitals’ telephone
switchboard (02890 240503) who will then contact the appropriate staff.
For routine results: Only available by telephone between 2 p.m. and 4 p.m.
Test results are available on the laboratory computer system which may be
accessed from designated VDUs.
Please avoid telephoning wherever possible. Non-urgent telephone calls create a
significant workload and cause unnecessary delay in processing samples.
Regional Immunology Laboratory
02890 632689
Internal
RGH ext 32689
Fax
02890 632622
P a g e | 141
For clinical advice, please contact:
Dr JDM Edgar (Consultant Immunologist)
02890 636649 or RGH ext 36649
david.edgar@belfasttrust.hscni.net
02890 632663 or RGH ext 32663
Dr L Devlin (Consultant Immunologist)
02890 631803 or RVH ext 31803
lisa.devlin@belfasttrust.hscni.net
02890 633853 or RVH 33853
Dr H Shendi (Consultant Immunologist)
02890 630002 or RVH ext 30002
Hiba.shendi@belfasttrust.hscni.net
Dr Cathal Steele (Specialist Registrar)
02890 631804
Cathal.steele@belfasttrust.hscni.net
For laboratory advice, please contact:
Dr Lynn Maxwell (Clinical Scientist)
02890 632623 or RGH ext 32623
lynn.maxwell@belfasttrust.hscni.net
Mrs Gillian Foster
(Immunology Operational Manager)
02890 632623 or RGH ext 32623
gillian.blair@belfasttrust.hscni.net
Mr Sean Conlan
(Immunology Services Manager)
sean.conlan@belfasttrust.hscni.net
02890 632643 or RGH ext 32643
P a g e | 142
Test repertoire
We provide a comprehensive range of tests for the immunological investigation of
patients. Our aim is to provide the highest quality of service with prompt delivery of
accurate results, (backed up by specialist medical and scientific expertise). Where
specific tests are not available locally, we will refer samples on to colleagues in other
centres.
The department is happy to assist in the interpretation of patient's test results.
Interpretative comments will be added to reports where appropriate.
Comments on how our service could be improved are always welcomed.
A full list of tests offered is described in the following pages and includes type and
volume of specimen and, if appropriate, any special requirements. There is a brief
summary of the clinical application of each test which is intended to be helpful but is
not intended to replace discussion of individual patients. The final section is a
"Disease Index" which is intended to assist in the selection of the most appropriate
investigations.
Turnaround Time
Average test turn around times (TAT’s) in working days are quoted for the various
tests. The turn around times for tests referred to other centres are closely monitored
and are between 2 and 35 days.
Transportation of Samples
Samples from the Royal Hospitals can be sent by hospital courier or via the vacuum
tube system. Samples from other hospitals / GPs may be sent by the relevant
dispatch systems.
The following documents are available from the laboratory on request:
Health & Safety Rules for Porters & Couriers
Pneumatic Tube Transport of Clinical Specimens
Transport of Specimens to the Laboratory
Postal samples must be sent in accordance with the guidelines issued by the Post
Office in respect of postal transmission of pathological specimens.
For advice contact the laboratory.
P a g e | 143
High risk samples
The laboratory must be informed of any known or potential hazards associated with
samples sent.
Specimens of blood, serum and other body fluids from suspected carriers of
Category 3 pathogens (hepatitis B or C and HIV) must be clearly marked with hazard
stickers and enclosed in a sealed plastic bag. Request forms should also have a
hazard sticker.
For some types of sample, and specific categories of hazard, a restricted range of
services may be offered.
Unsuitable Samples
If a sample is unsuitable for testing a report will be sent to the requestor giving the
reason and requesting another sample.
Samples unsuitable for testing include pleural effusion for any test, inappropriate
presence or absence of anticoagulant, delayed cellular/ functional assay samples
and haemolysed and/or lipaemic samples.
Requesting additional examinations
Patient serum samples are held by the laboratory for approximately 3 weeks. During
this time the laboratory may be contacted for discussion on the appropriateness of
additional testing.
`Duplicate samples
For most tests, samples received within 7 days of a previous sample will not be
tested. The following comment will be printed on the report: Test Name – Not tested.
Sample already received within 7 days.
Exceptions to this rule are:
ANCA, Anti GBM Antibody: 1 day
Anti CCP Antibody: 3 months
Immunology request and report forms
The immunology request form has a light brown strip along the top, middle and
bottom. Supplies for locations outside of the Belfast Trust may be obtained by
telephoning the laboratory (02890 632689). Report forms are a buff colour.
The request form contains 4 sections, which refer to separate sections of the
laboratory: Autoimmune serology, Immunochemistry, Allergy and Cellular
immunology. Tests may be requested by ticking the appropriate box or writing the
P a g e | 144
test required in the space provided. Please note, separate blood samples and
request forms are needed for tests performed in separate sections of the
laboratory. Refer to the laboratory sections in this handbook for further details
of their individual sample requirements. When a clotted blood sample is
required, a yellow topped tube MUST be provided.
In order to maintain high standards of practice and patient safety, the Belfast Trust
Laboratories (BTL), will ensure that stringent minimum acceptance criteria are in
place for the receipt and identification of samples.
Attention to detail is essential to ensure that the right result is sent out on the right
patient. Specimens will not be accepted for analysis where the following essential
criterion in the Minimum Data Set is not met on the form or the specimen container.
Identification data
Sample
Request form
Patients full name
Essential
Essential
Date of birth or
Hospital number
Essential
Essential
Destination for report
Desirable
Date and time
Desirable
Desirable
Patients sex
Desirable
Desirable
Patients address
Desirable
Patients consultant
or GP
Desirable
Signature of
requesting clinician
Desirable
Clinical information
Desirable
Essential for Specific IgE,
ANA, ANCA and vaccine
requests.
Please note: if gender is not included on request form the laboratory will not be able
to provide gender-specific reference ranges or identify critical results. If the
Consultant is not listed the laboratory may not be able to telephone critical results for
outpatients.
P a g e | 145
Clinical information should be provided on the request form for all requests but is
essential for specific IgE, ANA & ANCA requests. For vaccine studies, it is essential
to state on the request form whether the sample is pre or post vaccination.
Samples with inadequate identifying information will be rejected.
Referral Tests
Specialised tests which are not available in the Belfast Trust may be sent to selected
referral laboratories for analysis by arrangement. The referral centre names are
provided with the laboratory reports. Further details are available upon request.
Data Protection
The legal requirement for the Trust and its staff to treat personal information
confidentially and hold it securely is set out in the Data Protection Act 1998.
The Belfast Health & Social Care Trust has the following document in place and it is
available via the BHSCT Intranet site or from the laboratory on request:
Reference TP026/08: Policy on the Data Protection Act 1998 and Protection
of Personal Information


Comments/Complaints
The Regional Immunology Service adheres to the Belfast Trust ‘Policy and
procedure for the management of complaints and compliments’. A copy is available
from the laboratory upon request. Comments or compliments should be directed to
the Immunology Laboratory Services Manager, Mr Sean Conlan by post, email or
telephone.
Tel: 02890 632643 or RGH ext 32643 sean.conlan@belfasttrust.hscni.net

P a g e | 146
AUTOIMMUNE SEROLOGY
Sample requirements: One yellow topped gel sample tube required (4.0ml).
Separate blood samples and request forms are needed for tests performed in
separate sections of the laboratory. Please provide clinical details on the
request form.
For specific disease associations please see antibody list below. All results
should be interpreted in the context of the patient’s clinical history.
If clinical advice regarding interpretation of results is required, please use the
contact details listed above.
CONNECTIVE TISSUE DISEASE
Antinuclear antibody specificities (ANA)
Antinuclear antibodies are associated with systemic lupus erythematosus,
connective tissue, rheumatological and autoimmune liver diseases. ANA may also
occur in a number of other conditions including juvenile chronic arthritis, Sjogren’s
syndrome, fibrosing alveolitis, autoimmune hepatitis, viral infections particularly EBV
and CMV and in drug reactions. The following antibodies are detected as part of the
ANA specificity test. TAT for all ANA tests below: 5 days
Anti ds DNA antibody
Anti-dsDNA antibodies are strongly suggestive of systemic lupus
erythematosus (SLE) although they are present in only 40-70% of patients
with this disease. Our present anti dsDNA profile includes two assays for
dsDNA. Positive samples are also tested for anti dsDNA antibody by crithidia,
see below
Anti dsDNA antibody: Results reported in IU/ml, 0-4 IU/ml negative, 5-9
equivocal range, ≥10 positive
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Anti chromatin antibody
These antibodies are against chromatin or nucleosomes and are found in
patients with SLE and drug induced lupus. They are also found in some
patients with Sjogrens syndrome and the antiphospholipid syndrome. Their
presence has also been linked to glomerulonephritis in SLE patients.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti ribosomal P antibody
These antibodies are found in 10-15% of patients with SLE, often in the
absence of antibodies to dsDNA, they can also be found in patients with
rheumatoid arthritis. Also associated with neuropsychiatric symptoms and
renal involvement. Antibody levels correlate with disease activity.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti R52 and anti Ro60 antibody
Anti Ro antibodies are found in patients with primary Sjogren’s syndrome,
subacute cutaneous lupus erythematosus (particularly photosensitivity),
neonatal lupus, congenital complete heart block in babies born to SLE
mothers (rare) and SLE with interstitial pneumonitis.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti La antibody
Anti La antibodies are detected in patients with primary Sjogren’s syndrome
and SLE.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
P a g e | 148
Anti Sm antibody
Anti Sm antibodies are very specific for a diagnosis of SLE, occurring in 2530% of Afro-Caribbean patients but in a much lower proportion of Caucasian
patients. Usually found in association with anti RNP antibody.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI positive
Anti Sm/RNP antibody
Antibodies to Sm/RNP occur in patients with SLE and mixed connective tissue
disease (MCTD).
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti Ribonucleoprotein (RNP) A and anti RNP 68 antibody
Antibodies to RNP (ribonucleoprotein) occur in patients with SLE and mixed
connective tissue disease (MCTD).
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti Scl-70 antibody
Anti Scl-70 (topoisomerase-1) antibodies are detected in 20-40% of patients
with progressive systemic sclerosis and 20% of patients with limited systemic
sclerosis. Such patients are more likely to develop facial skin rash and heart
involvement.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti Jo-1 antibody
Anti Jo-1 antibodies (histidyl tRNA synthetase antibodies) are found in 2040% of patients with aggressive polymyositis usually in association with
interstitial lung disease and arthralgia. Antibodies to other tRNA synthetases
are also associated with variant myositis syndromes.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
P a g e | 149
Anti centromere B antibody
These antibodies are found in patients with the limited cutaneous form of
systemic sclerosis and in the CREST variant (Calcinosis, Raynaud’s,
oEsophageal immotility, Sclerodactyly, Telangiectasia). Also found in up to
12% of patients with primary biliary cirrhosis, over half of such patients have
clinical signs of scleroderma.
Results reported as Antibody Index (AI), 0 - 0.9 AI negative, ≥1 AI
positive
Anti dsDNA antibody by crithidia. This assay is performed on samples which are
positive by multiplex assay (≥10 IU/ml). The assay has very high specificity but poor
sensitivity for SLE.
Anti dsDNA antibody (Crithidia): Results reported as positive or negative.
TAT: 14 days
Anti histone antibody
Anti histone antibodies are found in 18-50% of patients with SLE and in 95% of
patients with drug induced SLE. This assay is performed by the Supraregional
Protein Reference Laboratory, Sheffield. Results reported as units / ml, positive
>5 U/ml.
Anti phospholipid antibodies
Anti cardiolipin antibodies form part of a spectrum of anti phospholipid antibodies
including anti β2 glycoprotein 1 antibodies. They are found in patients with a variety
of diseases, such as infections, malignancies and autoimmune disease including the
anti-phospholipid syndrome (APS) which may be primary or occur as a secondary
complication of SLE. The major features of APS are thromboses (arterial or venous),
thrombocytopenia, recurrent spontaneous abortion, skin rash (livedo reticularis). The
diagnosis of antiphospholipid syndrome is based on the presence of clinical and
laboratory criteria, which include antiphospholipid antibody and lupus anticoagulant.
Antiphospholipid antibodies should be present on 2 or more occasions, at moderate
to high levels (>40 U/ml) at least 12 weeks apart and no more than 5 years prior to
clinical manifestations. A sample should also be sent to Haematology for coagulation
(Lupus anticoagulant) studies.
P a g e | 150
Anti IgG and IgM cardiolipin antibody
Reported as GPLU/ml for IgG ACA and MPLU/ml for IgM ACA: 0-19.9
Negative, 20-39.9 low positive, 40-79.9 medium positive, 80-160 high
positive
TAT: 5 days
Anti IgG and IgM β2 glycoprotein 1 antibody
Reported as U/ml for both IgG and IgM β2GPL1 antibodies: 0-19.9
Negative, 20-39.9 low positive, 40-79.9 medium positive, 80-160 high
positive
TAT: 5 days
Antibodies in patients with myositis / dermatomyositis.
These include antibodies to Jo-1, PL-7, PL-12, SRP, Ku, Mi-2 and PM/Scl.
This assay is performed by the Immunology Laboratory, Royal Free Hospital,
London..
Results for these antibodies are reported as positive or negative.
Antibodies in patients with systemic sclerosis
These include antibodies to RNA polymerases and fibrillarin. This assay is
performed by the Immunology Laboratory, Royal Free Hospital, London.
Results for these antibodies are reported as positive or negative.
RHEUMATIC DISEASE
Anti cyclic citrullinated peptide antibody (CCP)
Anti-cyclic citrullinated peptide (CCP) antibodies are present in early rheumatoid
arthritis (RA) and appear to be a marker of more erosive disease. The sensitivity of
anti-CCP is similar to that of RF but the test is more specific for RA.
Results reported in Units/ml, positive ≥3 U/ml. TAT: 5 days
P a g e | 151
GASTROINTESTINAL DISEASE
Coeliac disease antibody screen
Requests for coeliac disease antibodies are screened for IgA anti tissue
transglutaminase antibody, those positive are further tested for IgA anti endomysial
antibody. For diagnostic purposes these samples should be from patients taking a
gluten containing diet for at least 12 weeks. Tests for IgG endomysial antibodies are
performed on samples from patients with suspected coeliac disease and IgA
deficiency.
Anti tissue transglutaminase antibodies (TGA)
Tissue transglutaminase is the antigenic target for anti endomysial antibody
and these IgA class antibodies are tested in combination with anti endomysial
antibodies bringing the sensitivity for coeliac disease to nearly 100%.
Treatment with a gluten free diet leads to gradual disappearance of these
antibodies. They can also be used to monitor dietary compliance. Approx 10%
of coeliac patients are only positive for either endomysial or transglutaminase
antibodies.
Results reported as EliA U/ml. Negative <7 EliA U/ml, Equivocal 7-10 EliA
U/ml, Positive >10 EliA U/ml. TAT: 10 days
Anti endomysial antibodies (EMA)
These IgA class antibodies are very specific (90-100%) for coeliac disease
(CD) and dermatitis herpetiformis (DH). Treatment with a gluten free diet
leads to gradual disappearance of these antibodies. They can also be used to
monitor dietary compliance. IgG class anti endomysial antibodies may be
detected in IgA deficient patients with coeliac disease.
Results reported as positive or negative. TAT: 14 days
Interpretation of coeliac antibody results: IgA tissue transglutaminase (TTG) is a
useful screening test for coeliac disease, whereas IgA endomysial antibodies (EMA)
are more disease specific and will automatically be performed when the IgA TTG
level is >4. Both tests may become negative in patients with coeliac disease on a
gluten free diet. Duodenal biopsy remains the gold standard test for diagnosis.
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Anti gastric parietal cell (GPC) antibodies
Anti GPC antibodies are present in 95% of patients with pernicious anaemia in the
early stages and in patients with atrophic gastritis (type A). They are also associated
with other organ specific autoimmune diseases especially autoimmune thyroid
disease. Also found in the normal population (the incidence rising with increasing
age). Anti-intrinsic factor antibody is a better confirmatory test for pernicious
anaemia.
Results reported as a titre, positive titre >40. TAT: 10 days
Anti intrinsic factor antibodies (IFA)
Anti IFA antibodies are highly specific for pernicious anaemia and are found in up to
75% of patients. Highly specific if found in combination with gastric parietal cell
antibody. Anti-intrinsic factor antibody may be detected before anaemia develops.
Results reported in units/ml, Negative <6 U/ml, positive ≥6 U/ml.
TAT: 14 days
AUTOIMMUNE LIVER DISEASE
Antinuclear antibody
Please request ANA as a separate test and provide clinical details on the request
form.
Liver associated antibodies (the following three autoantibodies are detected as
part of the liver associated autoantibody screen)
Anti smooth muscle antibody
These antibodies can occur in high titres in patients with autoimmune
hepatitis. Low titre antibodies may be detected after infection.
Results reported as titre, positive >40. TAT: 10 days
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Anti mitochondrial antibody (M2)
Anti M2 mitochondrial antibodies are detected at high titre in 95% of patients
with primary biliary cirrhosis. They can also be found in patients (usually lower
titres) with chronic active hepatitis, autoimmune thyroiditis and Sjogrens
syndrome.
Other subtypes of mitochondrial antibodies have been described such as M1:
associated with positive syphilis serology and M5: associated with SLE and
the antiphospholipid syndrome.
Results reported as a titre, positive >40. TAT: 10 days
Anti liver kidney antibodies (LKM)
Anti LKM-1 antibodies are associated in patients with type 2a and 2b
autoimmune hepatitis. This is the most common form of autoimmune hepatitis
in childhood and has a particularly poor prognosis and can be associated with
hepatitis C infection. Anti LKM-2 antibody is associated with drug induced
hepatitis and LKM-3 antibody is associated with hepatitis D infection.
Results reported as titre, positive >40. TAT: 10 days
Anti M2, anti Liver cytosol-1 (LC-1) and soluble liver antigen (SLA) antibodies
These antibodies are found in patients with primary biliary cirrhosis and autoimmune
hepatitis 1and 2.
This assay is performed by the Immunology Laboratory, King’s College Hospital,
London.
Results for these types of antibody are reported as positive or negative.
ENDOCRINE DISEASE
Anti adrenal antibodies
These antibodies are detected in 60-70% of patients with idiopathic Addison’s
disease.
Results reported as negative or positive (with titre). TAT: 14 days
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Anti islet cell antibodies
These antibodies are found early in the course of type I diabetes mellitus, gradually
disappear with time. Not found in type II diabetes mellitus.
Results reported as negative or positive (with titre). TAT: 14 days
Anti glutamic acid decarboxylase (GAD) antibodies
These antibodies are found in >60% of patients with the stiff man syndrome (high
titre) and also in patients with type 1 diabetes mellitus. These assays are performed
by The Institute of Molecular Science, John Radcliffe Hospital, Oxford.
Results reported as U/ml, normal range 0-5 U/ml.
Anti ovary/testes antibodies
A number of antibodies react with various cell types within the ovary and testes.
Antibodies found in patients with Type 1 autoimmune polyendocrinopathy syndrome
and premature gonadal and ovarian failure.
Results reported as negative or positive (with titre). TAT: 14 days
Anti pituitary antibodies
These antibodies are found in patients with lymphocytic hypophysitis and
autoimmune hypopituitarism.
This assay is performed by the Doctors Laboratory, London.
NEUROLOGICAL DISEASE
Anti acetyl choline receptor antibody (AChR)
These antibodies are found in 85 – 90% of patients with myasthenia gravis. 10-15%
of patients are sero-negative.
Results reported as antibody concentration.
< 0.45 nmol/L: negative: ≥0.45 nmol/L: positive TAT: 21 days
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Anti ganglioside antibodies (GM1, GQ1b)
These antibodies are associated with a number of peripheral neuropathies. Anti GM1
antibodies are associated with Guillain Barré syndrome (GBS), chronic
demyelinating polyneuropathy and multifocal motor neuropathy. Anti GQ1b
antibodies are associated with Miller Fisher variant of GBS. These assays are
performed by The Institute of Molecular Science, John Radcliffe Hospital, Oxford.
Anti ganglioside GM1 antibody: Results reported in units, normal range 0-200.
Anti ganglioside GQ1b antibody. Results reported in units, normal range 0-25.
Anti muscle specific kinase antibody (MuSK)
These antibodies are found in approx 40% of patients with generalised myasthenia
gravis who are negative for anti AChR antibody. These antibodies are sent to The
Institute of Molecular Science, John Radcliffe Hospital, Oxford for confirmation.
Results are reported as positive or negative.
Anti paraneoplastic antibodies (neuronal nuclear and purkinje cell)
These antibodies are associated with paraneoplastic disorders with accompanying
carcinomas. They include anti Yo (PCA), anti Hu (ANNA-1), anti Ri (ANNA-2)
antibodies, anti Ma, CV2/CRMP5 and amphiphysin antibodies. TAT: 10 days
These antibodies are screened in house and any suspected positives samples are
referred for further testing to confirm the antibody specificity. This is performed by
The Institute of Molecular Science, John Radcliffe Hospital, Oxford.
Results are reported as positive or negative.
Anti NMDA (N-methyl D-aspartate) receptor antibody
Described in patients with ovarian tumors and prominent psychiatric symptoms.
This assay is performed by The Institute of Molecular Science, John Radcliffe
Hospital, Oxford.
Results are reported as positive or negative
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Anti skeletal muscle antibody
These antibodies are present in some patients with myasthenia gravis and almost all
(80 – 100%) patients with thymomatous myasthenia gravis. They can also occur in
patients with hepatitis, acute viral infections and polymyositis.
Results reported as titre, positive >20. TAT: 14 days
Anti voltage gated calcium channel antibody (VGCC)
These antibodies are found in patients with the Lambert-Eaton myasthenic syndrome
(LEMS). This assay is performed by The Institute of Molecular Science, John
Radcliffe Hospital, Oxford.
Results reported in pM, positive >45pM.
Anti voltage gated potassium channel antibody (anti VGKC ab)
These antibodies are associated with acquired neuromyotonia. This assay is
performed by The Institute of Molecular Science, John Radcliffe Hospital, Oxford.
Results reported in pM, positive >100pM.
Anti Aquaporin 4 antibody
Antibodies found in 80% of patients with neuromyelitis optica (NMO) or Devic’s
disease and approx 50% of patients with longitudinally extensive transverse
myelitis.This assay is performed by The Institute of Molecular Science, John
Radcliffe Hospital, Oxford.
Results are reported as positive or negative
Anti basal ganglia antibody (ABGA)
These antibodies have been associated with Sydenham’s chorea, tic disorders and
encephalitis lethargic like syndrome, all associated with streptococcal infections. This
assay is performed by The Neuroimmunology and CSF laboratory, Institute of
Neurology, Queens Square, London .
Results are reported as positive or negative
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Beta interferon neutralizing antibodies
This assay is performed by The Neuroimmunology and CSF laboratory, Institute of
Neurology, Queens Square, London.
Results are reported as positive or negative
RENAL DISEASE ASSOCIATED ANTIBODIES
Anti neutrophil cytoplasmic antibodies (ANCA)
Indicated in the investigation of ANCA associated vasculitis. Main patterns
recognised, are cytoplasmic (C-ANCA) and perinuclear (P-ANCA). C-ANCA with
specificity for proteinase-3 (PR-3) has a high predictive value for active generalised
Wegener’s granulomatosis (WG) and can also be found in patients with microscopic
polyangiitis (MPA). P-ANCA with anti-myeloperoxidase (MPO-ANCA) specificity is
predictive for patients with active MPA and Churg Strauss syndrome (CSS), some
patients with WG also have this antibody.
P-ANCA with specificities other than MPO-ANCA occur in some patients with
inflammatory bowel disease, sclerosing cholangitis, rheumatoid arthritis, systemic
lupus erythematosus, chronic active hepatitis and other autoimmune diseases. In
such patients, ANCA levels are often low and of uncertain significance.
The assay will only be performed on patients with clinical features associated with
ANCA associated vasculitis (WG, MPA, CSS).
Results reported as a titre, positive titre >20.
All ANCA requests are tested for Anti PR3, anti MPO. See below.
Anti Proteinase-3 antibody (PR3)
Proteinase 3 (PR3) is the major target antigen for C-ANCA. The detection of anti
PR3-ANCA has a high predictive value for Wegener’s granulomatosis.
Results reported in Antibody Index units. Positive ≥1 AI
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Anti Myeloperoxidase antibody (MPO)
Myeloperoxidase is the target antigen for the majority of P-ANCA and is associated
with microscopic polyangiitis and Churg Strauss syndrome, but can also be found in
some patients with WG.
Results reported in Antibody Index units. Positive ≥1 AI
Anti glomerular basement membrane antibodies (GBM)
These antibodies are found in patients with Goodpasture’s syndrome (>90%
sensitivity)
Results reported in Antibody Index units. Positive ≥1 AI
TAT 2 days
Urgent requests can be tested within 2 hr of sample arriving at laboratory if
during normal working hours. Please contact the laboratory prior to sending.
C3 nephritic factor (C3 Nef)
These antibodies, to the alternative pathway C3 convertase, are found in patients
with membrano-proliferative glomerulonephritis (type II) and partial lipodystrophy.
These assays are performed by the Supraregional Protein Reference Laboratory,
Sheffield.
Results reported as detected or not detected. TAT: 35 days
OTHER AUTOANTIBODIES
Anti C1q antibodies
Antibodies to C1q may be associated with renal disease activity in patients with SLE.
High levels are found in patients with hypocomplementaemic urticarial vasculitis
syndrome (HUVS). These assays are performed by the Supraregional Protein
Reference Laboratory, Sheffield.
Results reported as ELISA U/ml, positive value >15 U/ml.
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Anti-cardiac muscle antibody
These antibodies are found in some patients with Dressler's syndrome and post
cardiotomy syndrome. This assay is performed by the Supraregional Protein
Reference Laboratory, Sheffield.
Results reported as negative or positive.
Anti-IgA antibodies
These antibodies may occur in patients with selective IgA deficiency. They can
cause blood product transfusion reactions. These assays are performed by NHS
Blood and Transplant Sheffield Centre, Sheffield.
Results reported as negative or positive with titre
Infliximab therapeutic drug monitoring
Infliximab (Remicade®), is a chimeric human-mouse monoclonal antibody directed
against tumour necrosis factor-alpha (TNF α ), approved for use in the treatment of
various chronic inflammatory diseases including rheumatoid arthritis, severe crohn’s
disease and ankylosing spondylitis. The drug is administered as an infusion with a
dosing interval ranging from 2 to 16 weeks.
Sample requirements: 0.5 mL of serum can be used for both infliximab drug levels
and anti-infliximab antibody analysis. The clotted blood sample should be received
by the laboratory within 4 hours of collection. Please contact the laboratory prior to
venepuncture as assay is by arrangement only.
To aid interpretation of results, it is essential that the following information is included
on the request form:
- Infusion dosing interval
- Number of infusions to date
- Reason for request, i.e., poor response
- Primary diagnosis
This assay is performed by the Clinical Biochemistry Department, City Hospitals,
Birmingham.
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Therapeutic ranges
It is suggested a cut-off for a therapeutic trough infliximab level of >1.0 μg/mL in a
patient on maintenance dose infusions.1
Reporting range
Our reporting range is 0.4 - 10.0μg/ml.
Other autoantibodies may be available on request: Please contact the laboratory.
IMMUNOCHEMISTRY
Sample requirements: One 4mL yellow topped gel sample tube is sufficient for
all immunochemistry measurements, unless otherwise stated below.
IgD
Sample requirements: one 4mL yellow topped gel sample tube.
Measurement of IgD is indicated in the investigation of hereditary periodic fever
syndromes.
These assays are performed by the Supraregional Protein Reference Laboratory,
Sheffield. Results reported as kU/L, normal range 2-100 kU/L.
IgE
See allergy section
Functional (specific) antibodies
IgG to tetanus toxoid.
Antibodies to pneumococcal specific antigens (PSSA) are available from Clinical
Immunology Laboratory Cambridge University Hospital.
Antibodies to meningococcal C, haemophilus and diphtheria are available from The
Meningococcal Reference Unit, Manchester.
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Functional antibody tests are of limited value, and are used mainly in the
investigation of primary immune deficiency. For advice, please contact immunology
medical staff.
Results are reported as: IgG TTox: mg/L, PSSA: ug/ml, MCA: rSBA titre,
HIB: ug/ml, DIP: IU/ml
TAT: 35 days.
CH50 and AH50 Functional Assays
Sample requirements: Samples must be received by the laboratory on the
same day the sample is taken.
Screening tests for classical (CH50) and alternate (AH50) complement activation
pathways are indicated in the investigation of suspected immunodeficiency
associated with recurrent pyogenic infections and atypical "immune complex
disorders". Values within the normal range indicate that the classical and alternate
pathway components are present. Quantitation of individual complement
components should be undertaken in samples with sub-normal CH50 and AH50
levels.
Normal ranges
CH50: 250-700 kU/L
APCH50: 66-129%. TAT: 10 days.
C1 esterase inhibitor (quantitative and functional)
Sample requirements: Samples must be received by the laboratory on the
same day that the sample is taken.
In type I hereditary angioedema (HAE) (85% of patients), low levels of C1 esterase
inhibitor (C1INH) are found by both the quantitative and functional assays. In type II
HAE (15% of patients) normal or raised levels of functionally inactive C1INH are
detected. Consequently, the functional C1INH assay is essential for this diagnosis.
Both types of HAE are associated with low or absent C4 levels during an attack.
Reduced levels of C1INH (quantitative and functional) and C1q are found in the rarer
acquired form of C1INH deficiency. This condition generally occurs secondary to
underlying disease, most frequently lymphoproliferative disorders.
Normal range:
C1INH (quantitative)
0.20 - 0.35 g/L
C1INH (functional)
70 -130%
TAT: quantitative – 10 days, functional – 14 days
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C1q
The primary indication for C1q measurement is in the differentiation of HAE (normal
C1q levels) from acquired C1 esterase deficiency (reduced C1q levels). Levels are
also decreased in conditions associated with immune complex mediated
complement activation.
These assays are performed by the Molecular Immunology Service, Cardiff and Vale
NHS Trust, Cardiff.
Normal range
50-250 mg/L. TAT: 35 days.
Mannose Binding Lectin (MBL)
Mannose binding lectin (MBL) plays an important role in the innate immune system
by facilitating complement activation. Measurement of MBL should be considered
when immunodeficiency associated with the complement system is suspected,
(recurrent infection, meningococcal disease).
The assay is performed by the Immunology Camelia Botnar Laboratories, Great
Ormond Street, London
Reference range:
<75 ng/ml correlates with homozygous variant alleles and non-functional MBL
which is associated with the greatest risk of infection.
75 – 399.9 ng/ml correlates with functional MBL deficiency associated with
increased risk of infection.
400 - 1300 ng/ml correlates with heterozygous variant alleles and may show
mild deficiency associated with some increased risk of infection.
> 1300 ng/ml correlates with wild type alleles showing no deficiency.
Individual complement components are available on request: Please contact the
laboratory.
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ALLERGY
Sample requirements: One 4.0mL yellow topped gel sample tube is sufficient
for all allergy testing, unless otherwise stated below.
Total IgE
Total serum IgE is usually elevated in patients with atopic disease. However levels
do not correlate with severity of disease and a raised IgE does not necessarily
indicate the presence of allergic disease. Other conditions where serum IgE levels
are raised include: parasitic diseases, some rare immunodeficiencies, atopic
eczema, eosinophilia, bronchopulmonary aspergillosis and in some lymphoid
malignancies.
Normal ranges
0-3 years
0-30 kU/L
4-7 years
0-70 kU/L
>7 years
0-120 kU/L
TAT: 5 days.
Allergen specific IgE
Allergen specific IgE testing is of value where skin testing is difficult to perform, or
contraindicated, ie.




in very young children.
in patients with severe/extensive eczema or dermographism.
in patients taking anti-histamines which cannot be stopped.
in patients in whom there is a significant risk of an anaphylactic reaction, the
use of allergen specific IgE testing must be carefully considered and is not a
substitute for careful clinical assessment.
High levels of specific IgE against a wide range of inhalant and food allergens are
frequently found in patients with atopic eczema. The clinical significance of such
sensitisation is often unclear.
Over 100 specific allergens are available for testing, however “screening“ for allergy
using allergen specific IgE is not usually helpful.
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If requesting allergen specific IgE testing, please provide as much clinical
information as possible, and which specific allergens are required.
The detection of allergen specific IgE in serum is not diagnostic of clinical allergy, nor
does the failure to detect allergen specific IgE exclude the diagnosis. Specific IgE
concentrations (Ku/L) do not correlate with clinical severity of allergic reactions.
TAT: 5 days.
Booklets are available on House Dust Mite Allergy and Peanut Allergy.
Copies may be obtained from the Immunology Secretaries in the Immunology Day
Centre. Ph 02890 635396 / 630003
Extrinsic allergic alveolitis screen
May be used in the investigation of patients with respiratory conditions in whom
hypersensitivity reactions to inhaled organic material is suspected. These conditions
are often associated with occupational exposure, for example, farmers’ lung
(thermophilic fungi) and bird fanciers’ lung (pigeons, caged birds).
Assays, which may be requested individually or as a screen include:
IgG antibodies to aspergillus fumigatus. Results in mg antigen/litre, normal
range <40.
IgG antibodies to micropolyspora faeni. Results in mg antigen/litre, normal
range <10.
IgG antibodies to pigeon protein. Results in mg antigen/litre, normal range <90.
IgG antibodies to budgerigar protein. Results in mg antigen/litre, normal range
<30.
TAT: 5 days.
Anaphylaxis
Sample requirements: Serial blood samples in 4mL yellow topped gel sample
tubes are required at the following times. 1st sample: as soon as resuscitation
has started, 2nd sample: 2 hours after reaction, 3rd sample: 24 hours after
reaction.
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Investigation of suspected anaphylactic reactions, in particular reactions occurring
during anaesthesia.
Investigations are recommended for patients with Grade II (cardiovascular reaction:
tachycardia, hypotension); Grade III (shock, life-threatening spasm of smooth
muscles); Grade IV (cardiac and/or respiratory arrest).
The following are measured: mast cell tryptase, IgE, allergen specific IgE (as
appropriate), C3, C4 (CLINICAL CHEMISTRY).
Notification of results will include interpretative comments and suggested
arrangements for follow up skin testing as appropriate.
TAT: 14 days.
Mast cell tryptase
Raised levels are detected during anaphylaxis. Timing of blood sample is critical as
maximum levels are observed within 3 hours post reaction. Elevated levels may also
be found in patients with mastocytosis.
Normal range:
2-14 ng/ml. TAT: 7 days.
CELLULAR IMMUNOLOGY
Investigation of the cellular immune system should only be undertaken after
discussion with medical staff. The appropriateness of testing and specimen
requirements will be advised. Samples should be received by the laboratory on the
same day as sample was taken.
Lymphocyte phenotyping
Sample requirements: 4ml EDTA blood sample.
Indicated in diagnosis and monitoring of immunodeficiency and in leukaemia
/lymphoma typing. Suspected cases of childhood T cell/combined immunodeficiency
(SCID) should be regarded as URGENT and the laboratory contacted as soon as
possible. Serial CD4 counts are of value in monitoring HIV disease, however
measurement of CD4 cells has no place in the diagnosis of HIV infection, until
serological status is established.
Requests for CD4 count as a “surrogate marker” of HIV infection will be refused.
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Lymphocyte subset panel: CD3 (T cell), CD4 (T helper), CD8 (T cytotoxic), CD19 (B
cell), CD16/56 (NK cell).
Markers of maturation, activation, monoclonality available by arrangement.
Results given as percentage and absolute counts (see appendix 1).
TAT: results within 24hr of sample receipt.
Lymphocyte Function
Sample requirements: Laboratory staff will advise of sample requirements and
pre-arrangement is essential.
Indicated in further definition of humoral and/or cellular immunodeficiency.
Proliferative responses to mitogens and/or specific mitogens are available.
Stimulants: Phytohaemagglutin (PHA), and Candida.
The assay is performed by the Immunology Camelia Botnar Laboratories, Great
Ormond Street, London
Neutrophil Function Tests
Sample requirements: Laboratory staff will advise sample requirements.
Indicated in investigation of recurrent skin infections, chronic gingivitis, recurrent
deep seated bacterial and fungal infections.
The following functional assays are available: Neutrophil Respiratory Oxidative
Burst.
Results given with reference to normal control value.
TAT: results within 24hr of sample receipt.
Cellular Analyses of Bronchoalveolar Lavage / Sputum
Sample requirements: Bronchoalveolar lavage washings and induced sputum
specimens.
Indicated in investigation of sarcoidosis, fibrosing alveolitis, and extrinsic allergic
alveolitis. Differential white cell counts are performed in the bronchoalveolar lavage/
sputum samples.
Results given as differential cell counts. TAT: 5 days.
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Disease index
Disease
Investigations
Addison’s disease
Anti-adrenal antibody
Allergy
IgE
allergen specific IgE
Anaphylaxis
mast cell tryptase
IgE
allergen specific IgE
C3, C4 (Clinical chemistry)
Angioedema
C1 esterase inhibitor
C1q
C3, C4 (Clinical chemistry)
Anti Phospholipid Syndrome (APS)
Anti-cardiolipin
glycoprotein I)
antibody
(anti-beta2
Lupus anticoagulant (Haematology)
Bullous Skin Disorders
Immunohistology (Tissue Pathology)
Skin reactive antibodies
Anti-endomysium antibody (in Dermatitis
Herpetiformis)
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Chronic Active Hepatitis
Anti-smooth muscle antibody
Anti-liver kidney microsomal antibody
Anti-mitochondrial antibody
Anti-nuclear antibody
Chronic Granulomatous Disease
Neutrophil function test
Chronic Lymphocytic Leukaemia
Immunoglobulins (Clinical chemistry)
Serum protein electrophoresis (Clinical
chemistry)
Lymphocyte phenotyping
Coeliac Disease
Anti-transglutaminase antibody
Anti-endomysial antibody
Congenital Heart Block
Anti-Ro antibody
Connective Tissue Diseases
Anti-nuclear antibody
Anti dsDNA antibody
Antibodies to ENA
CREST
Anti-centomere antibody
Cryoglobulinaemia
Cryoglobulins (Clinical chemistry)
C3, C4 (Clinical chemistry)
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Dermatitis Herpetiformis
Immunohistology (Tissue Pathology)
Anti transglutaminase antibody
Anti-endomysial antibody
Dermatomyositis
Anti-Jo-1 antibody
Diabetes
Anti-islet cell antibody
Anti GAD antibody
DLE
Anti-nuclear antibody
Dressler's Syndrome
Anti-cardiac muscle antibody
Extrinsic allergic alveolitis
IgG to aspergillus fumigatus.
IgG to micropolyspora faeni
IgG to avian proteins proteins (pigeon and
budgerigar)
Fibrosing Alveolitis
Anti-nuclear antibody
Glomerulonephritis
Anti-neutrophil cytoplasmic antibody
Anti-myeloperoxidase antibody
Anti-proteinase 3 antibody
Anti-GBM antibody
C3/C4 (Clinical chemistry)
Goodpasture's Syndrome
Anti-GBM antibody
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Graves Disease
Anti-thyroperoxidase antibody (Clinical
chemistry)
Anti thyroid hormone receptor antibody
(Clinical chemistry)
Guillain-Barre Syndrome
Anti-GM1 antibody
Anti-GQ1 antibody
Hashimoto’s Thyroiditis
Anti-thyroperoxidase antibody (Clinical
chemistry)
HIV Infection
Lymphocyte phenotyping
Immunodeficiency
Immunoglobulins (Clinical chemistry)
IgG subclasses (Clinical chemistry)
Functional antibodies
CH50
C3, C4 (Clinical chemistry)
CRP (Clinical chemistry)
Cellular investigations
Juvenile Chronic Arthritis
Rheumatoid factor (Clinical chemistry)
Anti-nuclear antibody
CRP (Clinical chemistry)
Leukaemia/Lymphoma
Cellular studies
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Lymphoproliferative disorders
Serum protein electrophoresis (Clinical
chemistry)
Immunoglobulins(Clinical chemistry)
Paraprotein detection and quantitation
(Clinical chemistry)
Cellular studies
Cryoglobulins (Clinical chemistry)
Mastocytosis
Mast cell tryptase
Membranoproliferative
Glomerulonephritis (MPGN)
C3 nephritic factor
Microscopic Polyangiitis
Anti-neutrophil cytoplasmic antibody
C3/C4 (Clinical chemistry)
Anti-myeloperoxidase antibody
Anti-proteinase 3 antibody
Mixed Connective Tissue Disease
(MCTD)
Anti-nuclear antibody
Anti-ENA antibody
Anti-RNP antibody
Multiple Sclerosis
CSF total protein (Clinical chemistry)
CSF IgG (Clinical chemistry)
IgG ratio and index (Clinical chemistry)
Oligoclonal bands (Clinical chemistry)
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Myasthenia Gravis
Anti-acetylcholine receptor antibody
Anti MuSK antibody
Anti skeletal muscle antibody
Non-Hodgkins Lymphoma
Immunoglobulins (Clinical chemistry)
Serum protein electrophoresis (Clinical
chemistry)
Cellular studies
Partial Lipodystrophy
C3 nephritic factor
C3, C4 (Clinical chemistry)
Pernicious Anaemia
Anti-gastric parietal cell antibody
Anti-intrinsic factor antibody
Pemphigus
Immunohistology (Tissue Pathology)
-Anti-intercellular substance / desmosome
antibodies
Pemphigoid
Immunohistology (Tissue Pathology)
-Anti-basement membrane zone antibodies
Polymyositis
Anti-Jo-1 antibody
Premature Ovarian Failure
Anti-adrenal antibody
Anti-steroid producing cell antibodies
Primary Biliary Cirrhosis
Anti-mitochondrial antibody (Anti M2)
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Progressive Systemic Sclerosis
Anti-nucleolar antibody
Anti-nuclear antibody
Anti-Scl-70 antibody
Raynaud's Phenomenon
Anti-centromere antibody
Rheumatoid Arthritis
Rheumatoid factor (Clinical chemistry)
CRP (Clinical Chemistry)
C3/C4 (Clinical chemistry)
Sjogren's Syndrome
Anti-nuclear antibody
Anti-Ro antibody
Anti-La antibody
Systemic Lupus Erythematosis
Anti-nuclear antibody
Anti-dsDNA antibody
Anti-ENA antibodies
Anti-cardiolipin antibody
C3/C4 (Clinical chemistry)
Vasculitis
Anti-neutrophil cytoplasmic antibody
Anti-myeloperoxidase antibody
Anti-proteinase 3 antibody
Immunohistology (Tissue Pathology)
CRP (Clinical chemistry)
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Wegener’s Granulomatosis
Anti-neutrophil cytoplasmic antibody
Anti-PR3-ANCA antibody
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Appendix 1 Lymphocyte Subset Reference Ranges
Lymphocyte
count
x 109/L
CD3
%
x 109/L
CD4
%
x 109/L
CD8
%
x 109/L
CD19
NK
%
x 109/L
%
x 109/L
0-2 months
1.97-7.98
69-93
1.38-7.13
48-75
1.17-5.62
13-33
0.27-1.86
4-26
0.1-1.45
2-14
0.006-0.43
2-4 months
3.89-10.75
55-79
2.53-6.78
35-62
1.39-5.21
14-30
0.65-2.45
14-39
0.75-3.5
3-14
0.19-0.99
4-11 months
4.18-12.21
59-80
2.89-7.99
38-61
1.79-5.14
14-34
0.95-2.97
16-36
0.86-3.77
3-13
0.18-1.58
11months- 2 yrs
3.46-11.62
50-78
2.2-8.19
26-53
1.09-4.55
16-39
0.75-3.75
16-33
0.70-2.71
5-19
0.18-1.58
2-3 yrs
2.05-6.14
52-78
1.35-4.09
26-50
0.7-2.44
11-38
0.43-1.74
17-34
0.52-1.78
5-19
0.16-1.17
3-4 yrs
1.73-5.22
44-79
0.89-3.65
25-53
0.49-1.72
13-37
0.27-1.54
17-35
0.39-1.54
4-22
0.12-0.64
> 4 yrs
1.56-4.57
54-79
1.05-3.03
28-49
0.55-1.72
17-32
0.33-1.31
9-32
0.2-1.14
6-25
0.14-1.03
Data from Chicago, USA
P a g e | 176
Microbiology Department
General Information
Department of Medical Microbiology
Kelvin Building, Royal Victoria Hospital, Grosvenor Road, Belfast, BT12 6BA
DX 3864NR
Laboratory Enquiries / Advice
028 9063 4140
Fax
(028) 9031 1416
Virology Specific Enquiries
028 9063 5242
028 9063 2662
Clinical Lead
Dr Anne Loughrey
028 9063 4112
Discipline Specific Manager (Acting)
Mr Mark Mc Gimpsey
028 9063 4125
Quality Lead
Mrs Karen Shields
028 9063 8275
Operational Manager Bacteriology
Mrs Nicola Wolsely
028 9063 2111
Operational Manager Specialist Services
Mr Mark McGimpsey
028 9063 8275
Operational Manager Molecular Services
Mr Frederick Mitchell
028 9063 3225
Public Health Laboratory
NIPHL
02890 263588
Belfast Health and Social Care Trust
Lisburn Road
Belfast
Northern Ireland
BT9 7AD
Infection Control Doctor
Dr Wesam Elbaz
Infection Control Nurses
028 9063 4020
028 9063 8160/8169
Duty Virologist
Dr Peter Coyle
(Clinical Virology advice Mon – Fri 9am –
5pm)
Dr Conall McCaughey
07889086946
Dr Susan Feeney
Dr Tanya Curran
Dr Alison Watt
Medical Microbiology Registrars
Office
028 9063 4117
P a g e | 177
(clinical advice Mon – Fri 9am – 5pm)
Pager (urgent only)
0425
Urgent Out of Hours
Microbiology Laboratory
028 9063 3607
07717731904
Virology
Contact Switchboard
Medical Microbiologist
Contact Switchboard
Laboratory Services
The laboratory offers consultant-lead scientific and clinical advice and interpretation on a
comprehensive range of tests for the microbiological investigation of patients.
Microbiology service comprises:







General Bacteriology
N. Ireland Mycobacterial Reference Laboratory
Mycology
Regional Virology Services
Serology
Molecular services (diagnostics, theranostics, epidemiology)
N. Ireland Public Health Laboratory (based on the City Hospital site)
Our aim is to provide the highest quality of service with prompt delivery of accurate
results, (backed up by specialist medical and scientific expertise). Where specific tests
are not available locally, they will be referred to colleagues in other centres.
Clinical Advice and Interpretation
Clinical advice or interpretation of results is available at all times. During working hours
contact the Medical Microbiology team as appropriate on the numbers available at the
start of the manual.
Out of hour’s Clinical advice:

Bacteriology
Contact on-call Medical Microbiology via switchboard.

Virology
Contact the Biomedical Scientist out of hours via switchboard in the first
instance. If necessary, out of hours requests and requests for medical advice
may be referred to a consultant virologist.
P a g e | 178
Request Forms
Requests should be written on the green and white Microbiology request form with
the exception of Regional Virology, Molecular and Serology, where dedicated forms
for specific services are available from the documents section of the laboratory
website: http://www.rvl-belfast.hscni.net. Requests can also be generated by order
com. Place the request form in the extra pocket of the plastic sample bag or attach it
outside with an elastic band. Do not staple the form to the bag.
Please avoid the terms “viral screening”, “routine virology”, “viral studies” or “viral
titres” as these terms are confusing and unhelpful. Instead, please provide brief
patient clinical details and duration of illness, which allows us to choose appropriate
tests.
Minimum Patient Identifiers
In order to fully identify a patient and send a full report back to the requesting source,
the request must contain the following information:
Essential
Sample
Request
Form
 Unique Identification Number
(E.g. H&C/ Hosp. / Client / WOC
No.)
 Forename
 Surname
 Date of Birth (dd/mm/yyyy)

Unique Identification Number
(E.g. H&C/ Hosp. /Client/WOC
No.)





HTA

Donor
Sample:
Request
form and
sample


Forename
Surname
Date of Birth (dd/mm/yyyy)
Test (s) Requested
Anatomical site and type of
specimen
Unique donor ID code (unique
patient identification number e.g.
H&C/Hosp. number)
Time
Place
Desirable
 Date & time




Destination for report
Sex
Name of Consultant or Gp
Date & time of sample collection
(which is sometimes essential)
 Patient’s address including
postcode
 MRSA/ESBL status etc. is
desirable for Bacteriology requests
 Relevant clinical information.
P a g e | 179
Please indicate if an unusual or fragile organism is suspected as the causal
agent, these may not be isolated by normal testing protocols and may require
special media, special isolation conditions and prolonged incubation.
Criteria for Rejection of Specimens
Specimens may be rejected immediately if:
1. Form Received with no specimen
2. Discrepancy between the patient details on the form and the specimen
3. No patient details on either the request form or the specimen
4. Specimens which have leaked and are insufficient for testing
5. Specimens deemed unsuitable for testing by BMS or clinical scientist at the
point of testing
6. Specimens accompanied by a request form with insufficient information to
send out a report or to determine which test is required.
When blood cultures are received that do not meet the minimum identifier set every
effort will be used to contact the ward/department by telephone to advise that the
sample is to be rejected and a repeat sample is necessary.
The laboratory may choose to process a sample that may otherwise have been
rejected.
In the case of CSF, tissues, bronchial washes and irreplaceable fluids for TB culture
and other such samples which cannot or are extremely difficult to repeat the
laboratory will contact the ward/department to clarify the patient identifier set. Note
will be taken as to whom communication has been made and included as a
laboratory comment on the report.
In other instances the final report will have a laboratory comment which indicates the
problem and if caution need be applied when interpreting the result.
P a g e | 180
Urgent Requests
It is essential that all request forms for emergency sample investigation are labeled
accordingly and prior arrangements made with the laboratory by phoning the
appropriate laboratory during normal working hours:
Department
Extension No
Working Hours
Bacteriology
33607
Mon – Fri 08:00 – 20:00
N. Ireland Mycobacterial
Reference Laboratory
35798
Mon – Fri 09:00 – 16:00
Mycology
34166
Mon – Fri 09:00 – 17:00
Serology
35242
Mon – Fri 09:00 – 17:00
Molecular
33225
Mon – Fri 09:00 – 17:00
For urgent samples out of hours please contact Biomedical Scientist on the
numbers listed at the start of the manual.
Requesting additional examinations
If requesting a further test, this should be requested as soon as possible after
receiving the original report and will be dependent on specimen retention, quality and
volume requirement issues. Additional requests will be either added to the original
request or a new request generated as appropriate.
Labelling High Risk Samples
For suspected or known Category 3 pathogens, hazard warning Category 3
pathogen labels should be affixed both to the container and the accompanying
request form. If there is any doubt as to whether a specimen is high risk, please
contact the Microbiology laboratory.
Hazard Group 3 is defined as a biological agent that may cause severe human
disease and presents a serious hazard to employees; it may present a risk of
spreading to the community, but there is usually effective prophylaxis or treatment
available.
NB: Hazard warning Category 3 pathogen labels should be affixed to ALL samples
taken from patients with pyrexia of unknown origin (PUO) following foreign travel.
P a g e | 181
Examples of Category 3 Pathogens
Bacteria:







Bacillus anthracis (Anthrax)
Brucella species
Escherichia coli, vero-cytotoxigenic strains (e.g: 0157: H7 and others)
Mycobacterium tuberculosis
Mycobacteria other than tuberculosis (MOTT)
Salmonella typhi
Salmonella paratyphi
Shigella dysenteriae (Type 1)
Fungi:




Blastomyces dermatitidis
Coccidioides immitis
Histoplasma species
Paracoccidioides brasiliensis
Penicillium marneffei
Viruses:-



All viral hepatitis (except Hepatitis A)
HIV
Severe Acute Respiratory Syndrome (SARS)
Prion Proteins:-




Transmissible spongiform encephalopathies (TSE) e.g: the agents of CreutzfeldtJacob disease (CJD): variant Creutzfeldt-Jacob disease (vCJD)
Fatal familial insomnia
Gerstman-Straussler-Scheinker syndrome
Kuru
Hazard Group 4 is defined as a biological agent that will cause severe human
disease and is a serious hazard to employees; it is likely to spread to the community,
and there is usually no effective prophylaxis or treatment available.
Please contact the Medical Microbiology team immediately if you suspect a
group 4 pathogen e.g. Lassa, Marburg, Ebola and Crimean. Under NO
circumstances should any samples be taken from such patients without prior
consultation with the Medical Microbiology team.
P a g e | 182
Packaging and Transportation of Samples
Advice on packaging samples for transport to the Laboratory
There is a responsibility for the Laboratories to give guidance to service users on the
packaging and transport of diagnostic specimens being shipped to the laboratory.
This is discharged through the issuing of the following instructions.
C-17 - Pneumatic tube transport of specimens
C-18 - Road transport of specimens
The manager of every Ward, Clinic, or GP which sends specimens to the Belfast
Trust Laboratories must read these and must ensure their unit complies with these
directions. The Carriage of Dangerous Goods Regulations (2011) and ADR (2011)
place a clear legal responsibility on the sender and any agent they use to transport
diagnostic specimens by road. Internal transfer to the laboratories by the pneumatic
tube system is subject to risk assessments made under the HSAW order and the
COSHH regs.
These instructions will be posted on the Laboratory website once it is operational,
they were considered too large to include in the User Manual so they can in the
interim be obtained on request by e-mail from our Dangerous Goods Safety Advisor
(DGSA) ian.doherty@belfasttrust.hscni.net
Transportation of Samples
There is a legal responsibility and a duty of care on anyone who dispatches clinical
material (diagnostic specimens) to the Belfast Trust Laboratories. The legal
responsibility is to ensure that if required, the specimens are packaged and labelled
in compliance with the relevant road transport regulations (ADR/CDG). There is a
further legal responsibility under COSHH regulations, since clinical materials may
contain infectious agents, to ensure that the materials do not leak or injure anyone
involved in the transportation or the wider public and environment.
The duty of care (to the patient) is to ensure that the transport conditions do not
damage the material being sent for testing or otherwise interfere with the validity of
the test results, and to ensure the specimen reaches the laboratory in good condition
within an appropriate time frame for good clinical management of the case.
Specimens should be clearly labelled and dated. Place all specimens in leak proof
containers in sealed plastic bags.
NB: Category 3 samples should be double bagged and clearly identified.
P a g e | 183



Samples from the Royal Group of Hospitals can be sent via the vacuum tube
system (except cat 3 samples) or by hospital courier.
Samples from other hospitals / GPs may be sent by the relevant dispatch
systems.
Postal samples must be sent in accordance with the guidelines issued by the
Post Office in respect of postal transmission of pathological specimens.
Any referring Unit, Hospital, Clinic, GP Practice or Trust transporting specimens by
road (which includes postal services) should take professional advice and guidance
on the packaging and labelling of any materials they hand over for transportation.
The laboratories are not responsible for nor do they have any managerial control
over the transportation of specimens between the shipper and the destination. The
strong recommendation by Belfast Trust Microbiology is that all patient Clinical
Specimens should be considered as potentially infectious and must therefore be
categorised at the very minimum as UN3373 Biological Substance Category B and
be packed and labelled according to Packing Instruction P650 in the ADR/CDG
regulations. (The packaging standard comprises 3 layers, two leak proof layers, and
a third outer rigid layer which provides protection against impact.)
If fully compliant with P650 then the package, the transport vehicle and the driver are
not subject to further specific requirements under ADR.
THIS EXEMPTION MUST ALWAYS BE USED
If the packaging is not P650 compliant then there is no exemption from the full
ADR/CDG regs, and the shipper and the driver will probably be found in breach of a
number of transport regulations and liable to prosecution.
All users’ will be contacted immediately if specimens are received that do not meet
with the stated packaging and transport requirements and informed as to how to
eliminate recurrence.
DX Courier Details
DX address DX 3864NR
Exchange: Belfast 14
P a g e | 184
Guidelines for proper specimen collection and factors which may affect the
quality of the results:





Confirm the identity of the patient either verbally and/or by hospital
identification wristband.
Complete the request form.
Check that the patient is appropriately prepared. This will obviously differ
depending on the nature of the sample(s) being taken.
Ensure the specimen(s) is collected correctly. It is important that the
laboratory receive good quality samples. Guidance on sample collection
especially those samples that are naturally eliminated from the body and
hence can often be collected by the patient can be found on the website
http://labtestsonline.org.uk/ (further information can also be found by
following the links in the specific sample types detailed below).
Collect specimen before administering antimicrobial agents when possible.

Check that the sample container is labeled correctly. Use sterile containers
and aseptic technique to collect specimens to prevent introduction of microorganisms during invasive procedures. Only laboratory approved, CE marked,
in vitro devices IVDs, must be used as primary specimen containers, no
substitutes or improvised containers.

Collect an adequate amount of specimen. Inadequate amounts of specimen
may yield false-negative results. If multiple samples are collected at the same
time ensure there is no interchange of samples.
Specimens obtained using needle aspiration should be transferred to a sterile
container and transported to the laboratory as soon as possible. If there is
only a small volume of material in the syringe, add some sterile preservativefree saline, mix and transfer to a sterile container. Do not send needle or
syringe.
All materials used in specimen collection should be disposed of safely
according to documented protocols.
All high risk samples MUST be identified to facilitate the correct processing of
such samples by laboratory personnel.
Any spillages or breakages occurring during sample collection must be dealt
with correctly according to a documented procedure.





All container tops must be firmly and properly closed, leakage adversely
affects not only that specimen but other specimens sharing the transit

The date and time of collection should be clearly stated (24 hr clock)
Fragile organisms may be affected by a delay in transport.
P a g e | 185

Specimens must be kept in a cool room awaiting dispatch, not in the sunlight
or near a radiator. Ensure the samples are stored under the appropriate
storage conditions for the investigation required.

Transit to the laboratory should be prompt and specimens must not be left in
uncontrolled vehicles (hot/cold) for any prolonged period.

If processing is delayed, refrigeration is preferable to storage at ambient
temperature. Delays of over 48h are undesirable.

Indicate if an unusual or fragile organism is suspected as the causal agent,
these may not be isolated by normal testing protocols & may require special
media, special isolation conditions & prolonged incubation.

To minimise the risk and ensure the safety of the specimen collector, carrier,
general public and the receiving laboratory, it is important that care is taken
when collecting and handling clinical samples to ensure that the risk of
infection to staff is kept to an absolute minimum. Therefore:
o Samples must always be carried in closed sealed plastic bags placed in
closed sealed boxes.
o Safe working practices shall be observed at all times.
o All clinical samples must be placed inside a sealed plastic bag.
o Should any urgent samples be sent outside of normal laboratory hours
(0900 – 1730) they will be transported in sealed plastic bags.
o Samples must never be carried unprotected in the open hand or given to
other members of staff in this way.
o Patient confidentiality must be preserved by the use of envelopes or
opaque plastic bags.
o Samples must always be carried in closed boxes which are clearly marked
with a BIOHAZARD label. Samples must never be thrown into a large
plastic bag and transported in this manner.
o The containment of samples within motor vehicles, used to transport
samples, must be such as to restrain, retain and protect the contents in the
event of an accident.
Unsuitable Samples If a sample is unsuitable for testing a report will be sent to the
requestor giving the reason and requesting another sample.
P a g e | 186
Results
Please avoid phoning whenever possible. The issuing of results of a non-urgent
nature over the phone is discouraged and must be kept to an essential minimum in
the interests of safety as verbal reports may lead to transcription errors.
Reports for both routine and emergency requests will be on labcentre and can be
viewed in your ward/practice as soon as they are validated by laboratory personnel.
Please make use of this facility.
If a phone enquiry is absolutely necessary, consult the laboratory computer system
to obtain the on-screen laboratory test request number, if this is available. This is in
order to validate that you are authorised to receive the confidential laboratory report
in question and will assist laboratory staff in dealing with your enquiry more
efficiently. All laboratory results should be interpreted in conjunction with the clinical
state of the patient. If inappropriate results are received, please contact the
laboratory and/or repeat the specimen.
Further Information
The laboratory has documented policies on Data Protection, Protection of Personal
Information and Complaints and Compliments available through the trust website
http://www.belfasttrust.hscni.net/services/Laboratory-MortuaryServices.htm.
Enquires and concerns can also be raised through the professional lead and
microbiology service manager.
P a g e | 187
Repertoire:
Please Note: Tests and specimen types listed below are for guidance only. For tests not listed below, or specimen types not listed within a particular test
please contact the laboratory to discuss clinical requirements.
Bacteriology Laboratory
Test
Sample Type
Container
Further information
TAT (Working Days)
Antibiotic Assay
5mls Clotted blood sample
red top bottle
60Gentamycin, Vancomycin, Amikacin, Tobramycin,
Teicoplanin Performed in Biochemistry Laboratory
Other Antibiotics Sent to reference laboratory
See Biochemistry
user manual
Blood culture
Adult – set of culture bottles
Aerobic –
Green top
Blood cultures should be taken aseptically according to Trust policy.
“No growth” will be
reported in 5 days
8-10mls Blood in each
Anaerobic –
Purple top
Paediatric – single bottle 1-3ml Blood
NB: In cases of endocarditis a maximal
volume of blood should be added
Blood Culture bottles are available from
microbiology specimen reception (Tel.
02890633507)
A maximum delay of 4 hours has been stipulated by National
UK Standards, PHE between inoculation of blood culture
bottles at the bedside and incubation of these in the lab.
Blood culture bottles should be inoculated before other containers
Yellow top
Repeated cultures increase sensitivity and help to distinguish
contaminants from clinically relevant organisms. Three sets, taken
not less than one hour apart, will give a success rate of 99%.
In cases of suspected intravascular catheter related sepsis, separate
blood cultures should be taken from the various lines or line ports.
An additional blood culture from a peripheral venepuncture should
be taken also.
Results of possible
pathogens will be
telephoned by
medical staff as a
priority.
P a g e | 188
Test
Sample Type
Container
Further information
TAT (Working Days)
Blood Cultures must NOT be refridgerated.
CSF
Up to 2mls CSF should be collected using
an aseptic technique.
Sterile
universal
container
CSF should be received into the laboratory within 1 hr. of being
taken.
Microscopy/cell
count – Same Day
In cases of suspected meningitis, the following should also be
considered:
Culture – Negative
& preliminary
results available
after 2 days


Ear
Swab
Blue cap
swab
Eye
Swab
Blue cap
swab
Corneal scraping should be accompanied
by a conjunctival swab
Glass slide
and brain
heart infusion
container
Blood culture
EDTA blood sample 2.5mls for meningococcal and
pneumococcal PCR
 Cryptococcus CSF/blood antigen if relevant
 Throat swab for meningococcal culture
 Haemorrhagic skin rash swab and glass slide
(pinch the lesion to exclude circulating blood and puncture with a
sterile needle. Squeeze a drop of fluid and smear on a slide.
Before sampling, remove debris with sterile saline under direct vision
Negative &
preliminary results
available after 2
days
-Conjunctiva
Eye
-Corneal scrape
The first corneal scraping should be
Negative &
preliminary results
available after 2
days

Microscopy and culture (bacterial and fungal) is routinely
performed on corneal scrapings.
Microscopy – Same
Day

If Acanthamoeba infection is suspected, please send a
punch biopsy or corneal scrape (without the blade) in
Culture – Negative
& preliminary
results available
P a g e | 189
Test
Sample Type
Container
Further information
approximately 200µl of sterile saline to the laboratory.
spread on a glass slide for gram staining

The second scraping sample should be
added with the blade to a brain heart
infusion container [supplied by the lab]
Faeces
Diarrhoeal samples which flow to assume
the shape of the container

Blue top
universal
container
with plastic
spoon
Aseptically collect 20 ml of pleural fluid.
If TB is suspected, a larger volume is
required (up to 250 ml if possible)
If a small volume of material is obtained
in the syringe, add some sterile


If enteric fever is suspected, please send
a blood sample for serology. Please refer
to the serology section.
Fluid [pleural]



Sterile
universal
container or
any large
sterile
container.


If Chlamydial or viral infection is suspected, please send an
additional conjunctival swab to the Regional Virus
Laboratory.
If mycobacterial infection is suspected, an extra slide should
be sent for ZN stain and a further sample in a sterile
universal container mixed with one ml of sterile preservativefree saline
TAT (Working Days)
after 2 days
Suspected
Acanthamoeba
specimens sent to
reference lab.
Diarrhoeal Samples are routinely tested for salmonella,
shigella, campylobacter, E. coli O157, cryptosporidium and
giardia.
Specimens from ICU patients and patients above 65 years
will be routinely tested for C.difficile . Otherwise, this needs
to be specifically requested.
Faeces will only be tested for other parasites on request and
in the presence of appropriate history ie foreign travel.
If you suspect a viral etiology please refer to virology section
http://labtestsonline.org.uk/understanding/analytes/ocp/tab/s
ample
Culture - Negative
& preliminary
results available
after 2 days
Microscopy (Gram and ZN stains) and culture (bacterial and
mycobacterial) is performed routinely. Please refer to the
mycobacterial section
Cell count will be performed on request only.
Microscopy/cell
count – Same Day
C difficle – 1 day
Parasitology – 1
Week
Culture – Negative
& preliminary
results available
P a g e | 190
Test
Sample Type
Container
Further information
preservative-free saline, mix
TAT (Working Days)
after 2 days
and transfer to a sterile container.
Fluid [joint]
Fluid should be collected under complete
aseptic conditions If only a small volume
of material is obtained, add some sterile
preservative-free saline, mix and transfer
to a sterile container.
Sterile
universal
container
only.
For cell count
please send
EDTA
sample







Helicobacter
Gastric Biopsy
a vial of
Dents media

Pus (exudates)
Any volume of pus is preferable to a swab
Sterile
universal


Crystal analysis will be performed on request only.
Cell count will be performed on request only.
If septic arthritis is suspected, a blood culture should be
sent.
If reactive arthritis is suspected, faecal culture may be
considered for salmonella, shigella, campylobacter and
yersinia; Paired sera for antibody testing is also required.
If rheumatic fever is suspected, a throat swab and a clotted
blood sample for antistreptolysin O titre are appropriate.
If a sexually transmitted aetiology is suspected, please refer
to the genital
swab section of user manual
Cell count will be performed on request only. If TB
peritonitis is suspected, ZN stain and TB culture should be
specifically requested. See mycobacterial section.
The test is performed by the Laboratory of Gastrointestinal
Pathogens (HPA Coilndale). The ref lab supply ‘Dents’ ,
which is a preservation media used for Helicobacter. We
now have a limited supply of this in the lab. Anyone
requesting this will now obtain a vial of Dents media and a
referral form from the lab. Return the biopsy in it and the
referral form for dispatch. It is important that this is
dispatched ASAP as samples over 96hrs old are not
suitable for culture. Consultants requesting this have been
advised to get samples to the lab no later than midday
Thursday.
Routine bacterial culture will be performed on all samples.
Gram stain will be performed on request only.
Microscopy/cell
count – Same Day
Culture – Negative
& preliminary
results available
after 2 days
15 days from
receipt in ref. lab.
Microscopy/cell
P a g e | 191
Test
Sample Type
Container
of pus.
container
Further information

If an unusual infection is suspected eg TB, this should be
highlighted and requested specifically.
Purulent and mucopurulent samples are
ideal
Salivary samples may be rejected
Wide neck
sputum
container




Samples should be taken prior to starting
antibiotic therapy.
Sputum production may be enhanced
with physiotherapy or saline inhalation.
If TB is suspected, three early morning
sputum samples on consecutive days
should be sent – please refer to the
mycobacterial section
count – Same Day
Culture – Negative
& preliminary
results available
after 2 days
If a small volume is obtained, add some
sterile preservative-free saline, mix and
transfer to a sterile container.
Sputum
TAT (Working Days)





Blood culture and pleural fluid culture may aid with the
diagnosis of pneumonia.
Urine for legionella and pneumococcal antigens should be
considered especially in community acquired pneumonia.
If Legionnaires` disease is suspected, sputum culture and
PCR should be requested.
ZN stain and mycobacterial culture will be performed on
request only.
In immunosupressed patients or patients with a history of
foreign travel where unusual mould infections are
suspected, fungal culture should also be specifically
requested.
Suspected ureaplasma/ mycoplasma hominis pneumonia in
ventilated neonates: send ET or tracheal secretions to
microbiology (0.5mls).
Inform microbiology registrar prior to sending secretions so
that laboratory is aware that sample is being sent.
Cystic Fibrosis: The form must be clearly marked “Cystic
Fibrosis” or “CF” so that the sample can be processed
appropriately.
http://labtestsonline.org.uk/understanding/analytes/sputumculture/tab/sample
Negative &
preliminary results
available after 2
days
P a g e | 192
Test
Sample Type
Container
Further information
TAT (Working Days)
Swabs
If possible the sample is taken after
physiotherapy. The swab is rubbed over
the high pharyngeal area after the patient
has been asked to cough.
Blue cap
clear
transwab
These swabs are from Cystic Fibrosis patients unable to produce a
sputum sample.
Swabs will be cultured for typical CF pathogens.
The form must be clearly marked “Cystic Fibrosis” or “CF” so that
the sample can be processed appropriately.
Negative &
preliminary results
available after 2
days
Females
Blue cap
clear
transwab
[Cough Swab]
(Cystic
Fibrosis)
Swabs
[genital tract]
High Vaginal swab
Cervical Swab
Urethral Swab
For investigation, please refer to the
Regional Virus Laboratory Section 5.
[MRSA, VRE, CRE
screening]
-
Nasal swab
Perineum swab [groin]
All wounds
Aseptic catheter sample of urine if
catheterized
Vascular access site if signs of
infection are present
Faeces (VRE only)
Routine bacterial culture will be performed on high vaginal
swabs.
Gonococcal culture will be performed on request only.
Clue cells will only be tested for on request, or if there is an
appropriate history of bacterial vaginosis
Trichomonas testing will be performed on request only.
Negative &
preliminary results
available after 2
days
NB. When taking a genital tract specimen which may have particular
legal significance, you may need to contact a genito-urinary
physician for advice.
[Chlamydia]
Swabs



Males
Swabs

Blue cap
clear
transwab
Swabs from other sites are not helpful for screening and will be
rejected.
NB. The swab should be moistened with sterile saline 0.9% solution
before use.
A throat swab is not required for routine MRSA screening and
therefore is not normally processed in the laboratory.
If a patient is to receive treatment in another hospital and a throat
swab is requested by that establishment, it is imperative that the
Negative &
preliminary results
available after 2
days
P a g e | 193
Test
Sample Type
Container
Further information
TAT (Working Days)
requirement and reasons for this swab are made clear on the
request form, otherwise the throat swab will not be processed.
Swabs
Pus and tissue samples are superior to
swabs
Blue cap
clear
transwab
Before sampling, remove debris with sterile saline or water. Rub the
swab over the base of the lesion.
Negative &
preliminary results
available after 2
days
Blue cap
clear
transwab
It is important to swab the infected area.
[throat]
Rub the swab over the tonsillar areas and
the posterior pharyngeal wall. Rotate the
swab to ensure that all the infected
mucosa is sampled
Negative &
preliminary results
available after 2
days
Tips
[intravascular]
Clean the skin with antiseptic solution
before withdrawal of the catheter.
Sterile
universal
container
[skin and soft
tissue]
Swabs
Tissue
Sterile
universal
container
For small samples, add a minimal volume
of sterile preservative-free saline to avoid
dryness of the sample.
Negative &
preliminary results
available after 2
days
Intravascular line tips should be accompanied by peripheral blood
cultures
Send a 5cm length of tip
Tips [others]
Swabs will be cultured for Haemolytic Streptococci and relevant
Corynebacterium species.
Specimens will be examined for Vincent`s organisms if clinical
history is suggestive.
Intravascular line tips should only be sent if line related sepsis is
suspected. They should not be sent for routine culture.
Sterile
container
-
Urinary catheter tip is not an appropriate sample and will be
rejected.
Aspirated fluid is superior to drain tips.
Samples should ideally be sent prior to starting antimicrobial
chemotherapy
Negative &
preliminary results
available after 2
days
Microscopy – Same
Day
Culture–Negative &
preliminary
P a g e | 194
Test
Sample Type
Container
Further information
TAT (Working Days)
available after 2
days
Tissue [bone]
Urine
Five separate samples should be taken
intra-operatively and put in five separate
containers.
Sterile saline
and beads
container
Samples should ideally be sent prior to starting antimicrobial
chemotherapy
 Blood culture may aid with the diagnosis, particularly in
acute presentations.
Mycobacterial culture will be performed only on request.
Microscopy– Same
Day
Mid-stream specimen of urine (MSSU)
Monovette
container
(yellow cap)
All urines undergo screening by automated microscopy which counts
cells and bacteria. Only those specimens reaching a certain
threshold number of cells and bacteria undergo culture.
Microscopy – Same
Day
Monovette
with Boric
acid
container
(green cap) if
sample is not
transported
immediately
Specific patient groups where a false negative screening result is
particularly high risk will undergo culture irrespective of the
screening result. These are: children <16 years, pregnant women,
urology patients, neutropenic patients, transplant recipients, and
patients undergoing repeat testing following a previous equivocal
culture result or for persistent symptoms.
[Cleanse the genital area with soap and
water before micturition prior to collecting
an MSSU]
Catheter specimen of urine (CSU)
[To collect a CSU, clamp the drainage
tubing below the sampling port and
aspirate sample with a sterile needle and
syringe. Unclamp.Do not collect from
drainage bag)
(must fill to
line)
Clean catch/pad specimen of urine
To ensure that all such patients’ urine specimens routinely
undergo culture please indicate clearly on the request form
when a patient belongs to one of these groups.
http://labtestsonline.org.uk/understanding/analytes/urine-
Culture – Negative
& preliminary
results available
after 2 days
Culture – Negative
& preliminary
results available
after 1 day
P a g e | 195
Test
Sample Type
Container
(paediatrics)
Urine
As above
Further information
TAT (Working Days)
culture/tab/sample
Sterile
container
Same Day
Legionella antigen
Streptococcus
pneumoniae
antigen
Mycobacteria Laboratory
Test
Sample Type
Container
Further information
TAT(Working Days)
Sputum
Wide neck
sputum container
Three fresh purulent samples should be collected at intervals of 824 hours, including one early morning sample.
Samples taken closer together may be combined into one request.
Samples without date and time information may also be combined.
Samples must be sent promptly to the lab.
Auramine Stain
Auramine Stain
Minimum 5ml
Culture
Bronchial washing/Broncho-alveolar
washing
1 Working Day of
receipt of sample
Plain sterile
container
Culture
Minimum 5ml
P a g e | 196
Test
Sample Type
Container
Further information
TAT(Working Days)
Sensitivities on all
Urine -The entire volume passed
Plain sterile
container
Three early morning samples on three consecutive days should be
collected.
Negative results
in 10weeks.
BORIC ACID
PRESERVATIVE
MUST NOT BE
USED
24hr collections of urine are not satisfactory.
plain sterile
container
Pleural and pericardial fluids are not very satisfactory samples due
to the low number of bacilli present. Pleural and pericardial biopsies
taken with the fluid give better results.
Swabs dipped in pus are rarely satisfactory and should only be
used when very small amounts of material are available.
MTBC primary
isolates.Sensitivities
on MOTT on
request.
Aspirated fluid and pus
Volume required, up to 250ml
Tissue
HISTOLOGICAL FIXITIVES MUST
NOT BE USED
Small biopsies may be sent in sterile
preservative-free saline to prevent
drying out.
CSF
Volume required,>6ml
For Neonates 2-4ml
Smaller volume will be tested down to a
minimum of 1.0ml, but the sensitivity of
the results will be low
(see Further Information)
Universal
containers,
sputum jars or
larger glass jars
may be used,
depending on
the size of
specimen.
Plain sterile
container
HISTOLOGICAL
FIXITIVES MUST
NOT BE USED
The British Infection Society guidelines for the diagnosis of
tuberculosis meningitis recommends approximately 10% of the total
CSF volume can be taken exclusively for mycobacterial testing.
Positive laboratory results are associated with large volume for
CSF submitted(>6.0ml)
Positive culture
results are
reported as they
arise. r PCR for
MTBC/Rif
resistance
detection on
direct positive
results within 1
working day.
MTBC
Sensitivities
14-21 days from
positive culture
identification.
P a g e | 197
Test
Sample Type
Container
Blood and bone marrow Maximum 5ml
Special culture
bottles are
available on
request from the
Microbiology lab.
Further information
TAT(Working Days)
A separate
sample of bone
marrow should
also be sent in a
plain sterile
container
Antibiotic Assay
(Rifampicin,
Ethambutol,
Isoniaizid
Pyrazinamide
Streptomycin)
Gastric washings Minimum 5ml
Plain sterile
container
Faeces
The HPA does
not recommend
culture of
faeces for
mycobacteria.
Contact NIMRL
Contact NIMRL
Blood
Blood collection
tubes (Grey cap,
Red cap and
Purple cap)
Collect samples early in the morning (before breakfast) on three
consecutive days.
Samples should be delivered to the TB lab within 4 hours or
neutralised with sterile 6.8ph phosphate buffer.(available from the
TB lab)
Please contact the TB lab for advice.
Sent to Reference Lab, Contact NIMRL for instructions on
testing protocols.
2-4 working days
This test is performed in areas of the hospital where prior
arrangements have been made. Otherwise it is only available on a
case by case basis following approval by the Consultant
Microbiologist. For approved areas please contact bacteriology
2 Weeks
Samples are
P a g e | 198
Test
Sample Type
Container
QuantiFERON
Further information
TAT(Working Days)
(028 90634281) for blood collection tubes and guidelines on
collection, storage and transportation of this test.
Samples are to be sent Mon-Wed only and by 16:00 hrs. on the day
of venepuncture. Otherwise for test approval in individual cases
please phone the duty micro SPR on 02890634139.
referred to the
National TB Ref
lab, London
where they are
processed on a
weekly batched
basis.
Mycology Reference Laboratory
Test
Isolation of
Sample type
Container
Further information
TAT(Working Days)
Skin
Skin lesions are
sampled by
Factors affecting the quality of results:
2 Weeks

Insufficient sample
P a g e | 199
Test
Sample type
Container
Further information
dermatophyte fungi
for the diagnosis of
dermatophyte
infections.
Hair
scraping with a
blunt scalpel and
collecting the
scales in folded
paper, which is
then folded
again and made
secure with a
paper clip, with
hairs and nails
also being
collected in this
manner.
MycoTans or
similar paper
packs designed
for this
 Formaldehyde or preservatives used
 Scrapings stuck to selotape
Identification, usually to species level. Yeasts are identified by a
combination of morphological and nutritional/enzymatic tests.
Moulds are usually identified on the basis of macroscopic and
microscopic morphology.
Fungi not considered to be clinically significant may not be
identified.
Nails
TAT(Working Days)
purpose may
also be used.
Cryptococcal
antigen.
CSF
Serum or CSF,
300μl minimum
or 2mls of
clotted blood.
4 Days
Isolation of yeasts
such as Candida
species for the
diagnosis of
Swabs, urine, Fluids, exudates
See General
bacteriology
section
3 Working Days
P a g e | 200
Test
Sample type
Container
Further information
TAT(Working Days)
Antifungal
sensitivity testing
Isolates of yeasts
Sabouraud’s
slope in a bijoux
or universal
Candida krusei is intrinsically resistant to fluconazole and is
therefore not tested against this antifungal. Sensitivity testing for
moulds is not generally indicated.
5 Days
Isolation of moulds
such as Aspergillus
species for the
diagnosis of
aspergillosis
Sputa and bronchoalveolar lavage
See General
bacteriology
section
1 Week
significant moulds
Fluids, exudates
See General
bacteriology
section
1 Week
Isolation of moulds
such as Sporothrix
schenkii for the
diagnosis of
mycetoma.
Biopsy and tissue
See General
bacteriology
section
4 Weeks
Antifungal drug
susceptibility
testing of moulds
Isolates of moulds
Candida Antigen
clotted blood
candidosis
Sent to reference lab.
BHSCT only
red top bottle
5-10mls
Antifungal Assays
(Flucytosine,
7 Days
clotted blood
Sent to reference lab.
7 Days
BHSCT only
red top bottle
Sent to reference lab
2 Days
P a g e | 201
Test
Sample type
Itraconazole,
Voriconazole)
5-10mls
Histoplasma
serology
Coccidioides
serology
clotted blood
Container
Further information
TAT(Working Days)
BHSCT only
red top bottle
5-10mls
Sent to reference lab.
14 Days
Travel history essential
BHSCT only
Paracoccidioides
serology
Blastomyces
serology
Aspergillus antigen
5.6 Serology
Test
Sample type
Container
Antistreptolysin O
5-10mls clotted blood
red top bottle
ANTHRAX:
Bacillus anthracis
5-10mls clotted blood
red top bottle
Sent to reference lab
21 Days
Bartonella
5-10mls clotted blood
red top bottle
Sent to reference lab.
21 Days
(Cat Sctatch)
Further information
TAT(Working Days)
3-5 Days
P a g e | 202
Test
Sample type
Container
Further information
TAT(Working Days)
B. pertussis IgG
5-10mls clotted blood
red top bottle
Sample should be taken > 3 weeks after onset for patients with a
history of prolonged cough (Sent to reference lab.)
21 Days
Refer to molecular test if <3 weeks post onset.
Borrelia burgdorferi
antibody) IgG and
IgM (Lyme
Disease)
5-10mls clotted blood
red top bottle
Sent to reference lab.
21 Days
Brucella IgG & IgM
ELISA
5-10mls clotted blood
red top bottle
C. diphtheria
5-10mls clotted blood
red top bottle
Toxigenic C.diphtheriae are very uncommon within the UK and are
almost always imported. A travel and immunisation history should
always be obtained from suspected cases of diphtheria.
(Sent to reference lab)
21 Days
Enteric
serodiagnosis
5-10mls clotted blood
red top bottle
Yersinia / Yersinia biotyping
21 Days
7-10 Days
E. coli including O157
Salmonella
Clostridium tetani
(Sent to reference lab)
L.pneumophila
clotted blood
Sg. 1-7 IgM ELISA
5-10mls
red top bottle
th
After about the 10 day post onset of disease, specific antibodies
are detectable in ELISA, however seroconversion can take up to 14
weeks (generally 3-6 weeks). Consequently early diagnosis cannot
be performed with serologic methods.
7-10 Days
P a g e | 203
Test
Sample type
Container
Further information
TAT(Working Days)
Leptospira IgM
ELISA
clotted blood
red top bottle
IgM antibodies can already be detected two days after the onset of
symptoms. These antibodies are detectable in all patients up to five
months after infection.
7-10 Days
red top bottle
Screen: Treponema pallidum Total Antibody
5 Days
5-10mls
Syphilis screening
and Confirmation
clotted blood
5-10mls
Confirmation of positive screen:



Toxoplasma gondii
IgM & IgG
clotted blood
red top bottle
5-10mls
Monospot
clotted blood
Treponema pallidum IgM
RPR
TPPA
Depending on clinical history and screening result samples may be
sent off to the Toxoplasma reference Laboratory for confirmation
and Dye Test
red top bottle
10-14 Days
.
3-5 Days
5-10mls
Rickettsia
clotted blood
red top bottle
5-10mls
Parasitic Diseases
clotted blood
5-10mls
Ehrlichia, Typhus Group, Spotted Fever Group
21 Days
(Sent to reference lab)
red top bottle
Amoebiasis; Babesia; Cysticercosis;
Fasciola; Filaria; Hydatid; Leishmania;
Malaria; Schistosomiasis Strongyloides; Toxocara; Trichinella;
21 Days
P a g e | 204
Test
Sample type
Container
Further information
TAT(Working Days)
Trypanosomal
+ other tropical diseases as requested
( Sent to reference lab)
Staphylococcal &
clotted blood
Streptococcal
serodiagnosis
5-10mls
TULARAEMIA:
Francisella
tularensis
clotted blood
red top bottle
Sent to reference lab
21 Days
red top bottle
Sent to reference lab
21 Days
5-10mls
Molecular Services
Test
Sample type
Container
Further information
TAT(Working Days)
Adenovirus
Respiratory samples
Sterile container.
PCR
5 Days
Nasopharyngeal secretions,
sputum,
tracheal
secretions,
bronchoalveolar lavage (BAL)
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
P a g e | 205
Test
Sample type
Container
Eye swab
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Urine 10-20ml
Blood 5ml EDTA
Sterile container
with no
preservatives
Further information
TAT(Working Days)
Do not use gel/charcoal swabs.
Purple top bottle
Adenovirus (faecal
group F)
Faeces
Blue top universal
container with
plastic spoon
PCR
5 Days
Arboviruses
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Astrovirus
Faeces
Blue top universal
container with
plastic spoon
PCR
5 Days
Bocavirus
Respiratory samples
Sterile container.
PCR
5 Days
Swabs in UTM,
Do not use gel/charcoal swabs.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
P a g e | 206
Test
Bordetella
pertussis
Sample type
Container
Further information
TAT(Working Days)
Throat/nasal swabs
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Respiratory samples
Sterile container.
PCR
5 Days
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Do not use gel/charcoal swabs.
It is only necessary to send one
specimen per patient as follows:
All genital swabs
and urine
specimens must be
sent in dedicated
specimen collection
kits which are
available from the
laboratories where
PCR
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Chlamydia
trachomatis
Females: Urine OR endocervical swab
OR vulvovaginal swab
NB. All genital swabs and urine specimens received for
Chlamydia trachomatis screening will be also tested for
Neisseria gonorrhoea.
5 Days
P a g e | 207
Test
Sample type
Males: Urine
Container
Further information
TAT(Working Days)
you deliver
microbiology
specimens to, i.e.
BCH, Mater, RVH
and Ulster
Hospitals.
A dedicated
specimen collection
kit may be used
when available. If
not available, a
swab in UTM or a
dry swab placed in
a sterile universal
container is
satisfactory.
Conjunctival swabs
Respiratory
Coronaviruses
Respiratory samples
Do not use gel/charcoal swabs.
Sterile container.
PCR.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
Swabs in UTM,
Do not use gel/charcoal swabs.
5 Days
P a g e | 208
Test
Sample type
Container
lavage (BAL)
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory
Throat/nasal swabs
Cytomegalovirus
(CMV)
Further information
TAT(Working Days)
Clotted blood 5ml
Red top bottle
Serology IgM
8 Days
Clotted blood 5ml
Red top bottle
Serology IgG immunity
3 Days
Clotted blood 5ml
Red top bottle
Gancyclovir resistance (Sent to reference laboratory)
21 Days
Clotted blood 5ml
Red top bottle
UL96 mutational analysis (Sent to reference laboratory)
21 Days
Clotted blood 5ml
Red top bottle
CMV IgG avidity (Sent to reference laboratory)
21 Days
EDTA blood 5ml
Purple top bottle.
PCR – Recommended specimen in adults.
5 Days
Urine 10-20ml
Sterile container
with no
preservatives.
PCR - Recommended specimen in infants with suspected
congenital CMV infection.
PCR
Fetal material
Colon biopsy
Sterile container
with no
preservatives.
PCR - Recommended specimen in suspected CMV colitis.
P a g e | 209
Test
Sample type
Container
Further information
TAT(Working Days)
Blue top universal
container
Dengue fever
Enteroviruses
(picornaviruses)
(inc Coxsackie &
parechoviruses)
Clotted blood 5 ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Faeces
Blue top universal
container with
plastic spoon.
PCR
5 Days
Throat swab
Clotted blood 5ml
Throat swabs in
UTM, eNat medium
or lysis buffer.
Red top bottle
Do not use gel/charcoal swabs.
Serology
8 Days
EDTA blood 5ml
Purple top bottle
PCR
5 Days
Clotted blood 5ml
Red top bottle
Immunofluorescence
30 Days
Clotted blood 5ml
Red top bottle
Serology IgM
8 Days
Clotted blood 5ml
Red top bottle
Serology IgG immunity
8 Days
Clotted blood 5ml
Red top bottle
PCR
5 Days
Fetal material
Sterile container
Red top bottle
Clotted blood 5ml
Sterile container.
CSF at least 0.5 ml
Epstein Barr virus
(EBV)
Nasopharyngeal
carcinoma Screen
Anti-EBV Viral
Capsid Antigen
Erythrovirus B19
(parvovirus B19)
P a g e | 210
Test
Sample type
Container
Further information
TAT(Working Days)
Haemorrhagic
Fever
Clotted blood 5ml
Yellow top bottle
Serology (Sent to reference laboratory)
21 Days
Hantavirus
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Hepatitis A virus
Clotted blood 5ml
Red top bottle
Serology IgM
3 Days
Clotted blood 5ml
Red top bottle
Serology IgG immunity
3 Days
Clotted blood 5ml
Red top bottle
Screening for infection (HBsAg)
3 Days
Clotted blood 5ml
Red top bottle
Markers
3 Days
Clotted blood 5ml
Red top bottle
Anti-HBsAg immunity
3 Days
Clotted blood 5ml or
Red top bottle or
Purple top bottle
Molecular quantitative (viral load)
14 Days
Clotted blood 5ml
Red top bottle
HBV DNA (occupational)
21 Days
Clotted blood 5ml
Red top bottle
Pre-core mutant analysis (Sent to reference laboratory)
30 Days
Clotted blood 5ml
Red top bottle
Lamivudine resistance analysis (Sent to reference laboratory)
30 Days
Clotted blood 5ml
Red top bottle
Serology screening for infection
3 Days
Clotted blood 5ml
Red top bottle
Antigen
8 Days
Clotted blood 5ml or
Red top bottle or
Purple top bottle
Molecular quantitative (viral load)
14 Days
Red top bottle or
Purple top bottle
Genotyping
21 Days
Hepatitis B virus
EDTA blood 5ml
Hepatitis C virus
EDTA blood 5ml
Clotted blood 5ml or
P a g e | 211
Test
Sample type
Container
Further information
TAT(Working Days)
EDTA blood 5ml
Hepatitis D virus
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Hepatitis E virus
Clotted blood 5ml
Red top bottle
Serology IgG and IgM (Sent to reference laboratory)
21 Days
Herpes simplex
virus (HSV 1 / HSV
2)
Clotted blood 5ml
Red top bottle
Serology IgM
8 Days
CSF at least 0.5ml
Sterile container
PCR
5 Days
Genital, skin, eye and swabs from other
sites.
Swabs in UTM or
eNat medium. If not
available, a dry
swab placed in a
sterile universal
container is
satisfactory.
Do not use gel/charcoal swabs.
Human Herpes 8
EDTA blood 5ml
Purple top bottle
PCR (Sent to reference laboratory)
21 Days
Human
Immunodeficiency
Virus (HIV)
Clotted blood 5ml
Red top bottle
Serology screen for infection
3 Days
EDTA blood 5ml x 2
Purple top bottle
Molecular quantitative (viral load) – sample must arrive at the
laboratory on the same day as it is taken.
14 Days
EDTA blood 5ml
Purple top bottle
Congenital transmission (Sent to reference laboratory)
21 Days
EDTA blood 5ml x 2
Purple top bottle
Genotypic resistance testing – sample must arrive at the
laboratory on the same day as it is taken.
30 Days
Clotted blood 5ml
Red top bottle
Serology
10 Days
Human T lymphotropic virus
(HTLV)
P a g e | 212
Test
Sample type
Container
Further information
TAT(Working Days)
Influenza viruses
Respiratory samples
Sterile container.
PCR
5 Days
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Legionella
pneumophila
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Respiratory samples
Sterile container.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Measles virus
NB. Recommended specimen is a combined nasal/throat swab.
However in ICU patients a lower respiratory tract specimen is
preferable if possible.
Do not use gel/charcoal swabs.
PCR
5 Days
Do not use gel/charcoal swabs.
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Respiratory samples
Sterile container
PCR
5 Days
P a g e | 213
Test
Sample type
Container
TAT(Working Days)
This test can only ensure reliable diagnosis if the sample is
taken within 10 days of onset. For information regarding
sampling beyond 10 days post onset, please contact the
laboratory.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Human
metapneumovirus
(hMPV)
Further information
NB. Recommended specimen type is a throat swab.
Do not use gel/charcoal swabs.
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Clotted blood 5ml
Red top bottle
Serology IgG (Sent to reference laboratory)
21 Days
Respiratory samples
Sterile container
PCR
5 Days
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container
Throat/nasal swabs
Do not use gel/charcoal swabs.
P a g e | 214
Test
Sample type
Container
Further information
TAT(Working Days)
Molluscum
contagiosum
Skin material
Scrape the
granulation tissue
underlying the skin
with a disposable
scalpel blade.
Transfer the
material to a clean
slide, air dry and
seal with a second
slide.
EM
10 Days
Mumps virus
Saliva / Urine
Sterile universal, no
preservatives.
PCR
5 Days
NB. Recommended specimen type is a parotid duct or buccal
membrane swab.
Do not use gel/charcoal swabs.
Swab
Mycoplasma
pneumoniae
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Clotted blood 5ml
Red top bottle
Serology IgG immunity (Sent to reference laboratory)
21 Days
Respiratory samples
Sterile container.
PCR
5 Days
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Do not use gel/charcoal swabs.
P a g e | 215
Test
Sample type
Container
Further information
TAT(Working Days)
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Neisseria
gonorrhoea
See Chlamydia trachomatis for details.
See
Chlamydia
trachomatis
for
details.

NB. If culture testing or antibiotic sensitivities are
required, a separate swab must be sent to bacteriology.
5 Days
Neisseria
meningitidis
Clotted blood 5ml
Red top bottle
PCR
5 Days
CSF at least 0.5ml
Sterile universal
NB. If culture testing or antibiotic sensitivities are required, a
separate specimen must be sent to bacteriology.
Respiratory samples
Sterile container.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Throat/nasal swabs
Do not use gel/charcoal swabs.
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
P a g e | 216
Test
Sample type
Container
Further information
TAT(Working Days)
swab placed in a
sterile universal
container is
satisfactory.
Norovirus
Faeces, vomit
Blue top universal
container
PCR
5 Days
Orf virus
Skin material
Scrape the
granulation tissue
underlying the skin
with a disposable
scalpel blade.
Transfer the
material to a clean
slide, air dry and
seal with a second
slide.
EM
10 Days
Parainfluenza
viruses
Respiratory samples
Sterile container.
PCR
5 Days
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Throat/nasal swabs
Do not use gel/charcoal swabs.
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
P a g e | 217
Test
Sample type
Container
Further information
TAT(Working Days)
PCR
5 Days
container is
satisfactory.
Pneumocystis
jiroveci (carinii)
Respiratory samples
Sterile container.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Do not use gel/charcoal swabs.
Pneumocystis
jiroveci (carinii)
Polyomavirus BK
Polyomavirus JC
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
EDTA blood 5ml
Purple top
Urine 10-20ml
Sterile universal, no
preservatives
CSF at least 0.5ml
Clotted blood 5ml
PCR
5 Days
Sterile universal
PCR (Sent to reference laboratory)
21 Days
Red top bottle
Serology IgM , Phase I & II immunofluorescence (Sent to
reference laboratory)
21 Days
P a g e | 218
Test
Sample type
Container
Further information
TAT(Working Days)
Q fever (Coxiella
burnetii)
Clotted blood 5ml
Red top bottle
PCR
5 Days
Sterile container.
Respiratory samples
Do not use gel/charcoal swabs.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Rabies virus
Clotted blood 5ml
Red top bottle
Serology IgG immunity (Sent to reference laboratory)
30 Days
Respiratory
syncytial virus
(RSV)
Respiratory samples
Sterile container.
PCR
5 Days
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Throat/nasal swabs
Do not use gel/charcoal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
P a g e | 219
Test
Sample type
Container
Further information
TAT(Working Days)
PCR
5 Days
sterile universal
container is
satisfactory.
Rhinovirus
Respiratory samples
Sterile container.
Nasopharyngeal secretions, sputum,
tracheal secretions, bronchoalveolar
lavage (BAL)
Do not use gel/charcoal swabs
Throat/nasal swabs
Swabs in UTM,
eNat medium or
lysis buffer.
Alternatively a dry
swab placed in a
sterile universal
container is
satisfactory.
Rickettsia
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Ross River virus
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Rotavirus
Faeces
Blue top universal
container
with
plastic spoon
PCR
5 Days
Rubella virus
Clotted blood 5ml
Red top bottle
Serology IgM
8 Days
P a g e | 220
Test
Sample type
Container
Further information
TAT(Working Days)
Clotted blood 5ml
Red top bottle
Serology IgG immunity
3 Days
Clotted blood 5ml
Red top bottle
PCR
5 Days
CSF at least 5 ml
Sterile universal
NB. If culture testing or antibiotic sensitivities are required, a
separate specimen must be sent to bacteriology.
Tick borne
encephalitis
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
Varicella zoster
virus (VZV)
Clotted blood 5ml
Red top bottle
Serology IgM
8 Days
Clotted blood 5ml
Red top bottle
Serology IgG immunity
5 Days
CSF at least 0.5ml
Sterile universal
PCR
5 Days
Streptococcus
pneumoniae
Genital, skin and swabs from other
sites.
Do not use gel/charcoal swabs.
Swabs in UTM or
eNat medium. If not
available, a dry
swab placed in a
sterile universal
container is
satisfactory.
Yellow fever
Clotted blood 5ml
Red top bottle
Serology (Sent to reference laboratory)
21 Days
P a g e | 221
Information regarding reference laboratories is available on request.
P a g e | 222
TISSUE PATHOLOGY
CONTACT US
We welcome comments from users about the content of the user manual to
enable us to improve the service or provide more appropriate information.
This may be done by contacting the Quality Lead indicated below.
In addition, specific groups of users are contacted annually by survey
questionnaire (Multidisciplinary Teams, General Practitioners and laboratories
that refer specimens to us) to establish the degree of user satisfaction
pertaining to the usefulness and timeliness or otherwise of our reports and to
solicit suggestions for improvement.
RGH Switchboard
028 9024 0503
BCH Switchboard
028 9032 9241
Tissue Pathology
Royal Victoria Hospital , Institute of Pathology, Grosvenor Road, Belfast, BT12 6AB
and
Belfast City Hospital, Corry Building, Lisburn Road, Belfast , BT9 7AB
Professional Lead
Dr Estelle Healy
028 9063 2545
Discipline Specific Manager
Mr Desy Smart
079 1788 0333
Quality Lead
Mr Ian Wallace
077 0451 0047
ianj.wallace@belfasttrust.hscni.net
Operational Manager – Surgical
Pathology
Mr Gerry Clarke
028 9063 2632
077 4769 8673
General Histopathology
078 2453 9469 (RGH)
Paediatric Pathology
028 9063 2170
Neuropathology
028 9063 2119
Paediatric Pathology Consultants
Dr Caroline Gannon
028 9063 8342
Dr Daniel Hurrell
028 9063 2369
Dr Brian Herron
078 0186 6678
P a g e | 223
Neuropathology Consultants
Dr Estelle Healy
077 7925 5420
Operational Manager – Diagnostic
Cytopathology
Ms Anna Patterson
028 9063 3832
078 2456 4169
Operational Manager – Cervical
Cytology
Ms Hilary Diamond
028 9504 8100
077 8043 1497
Operational Manager – Specialist
Services
Mr Gerry Clarke
028 9063 2632
077 4769 8673
Immunocytochemistry
028 9504 1248
Immunopathology
028 9063 2534
077 1780 5516
Electron Microscopy
028 9063 2670
Molecular Pathology
Operational Manager – Post-Mortem
Services
Dr Perry Maxwell
028 9097 2708
Prof Manuel Salto-Tellez
028 9097 2718
Mr John Murray
028 9063 2069
077 8607 6972
Adult Post-Mortem Consultant
Dr Brian Herron
028 9063 2613
Mortuary Office
028 9063 3679
Paediatric Post-Mortem Consultants
Dr Caroline Gannon
028 9063 8342
Dr Daniel Hurrell
028 9063 2369
Paediatric Office (PM Enquiries)
028 9063 3857/3858
Mortuary Mobile
079 7936 6041
Consultants (RGH)
077 7563 5395
P a g e | 224
TISSUE PATHOLOGY LABORATORY SERVICES
(Including opening times of the laboratory, out of hours service and
clinical advice & interpretation)
(CPA reference 1341 – accredited)
Tissue Pathology provides a diagnostic and reference service, not only to the
BHSCT, but also to the whole of Northern Ireland. Tissue Pathology samples
are usually submitted for the assessment of macroscopic, microscopic,
molecular and ultrastructural abnormalities to aid diagnosis, treatment
planning and management of disease states, including malignancy.
The following range of diagnostic specialties is available:









breast pathology
bone & soft tissue pathology
cardiovascular pathology
cervical cytology
diagnostic
cytopathology
(including
fine
needle
aspiration cytology)
dermatopathology
endocrine pathology
ENT pathology
gastrointestinal pathology










haematopathology
hepatopancreatobiliary
pathology
lung pathology
ophthalmic pathology
oral & dental pathology
paediatric pathology
renal pathology
ultrastructural pathology
neuropathology
urological pathology
These are underpinned by technical expertise in Immunocytochemistry,
Immunofluorescence, Molecular Pathology, Electron Microscopy and Image
Analysis.
Specialty
Surgical Pathology (General
Histopathology,
Neuropathology & Paediatric
Pathology)
Hours of Service
Out of Hours
(Monday-Friday)
(Saturday, Sunday, Bank Holidays &
urgent specimens)
8.30am – 5.00pm
General Histopathology – contact duty
consultant pathologist via BCH or RGH
switchboard (Lab Staff 07824539469)
BCH & RGH
Neuropathology – specify either Dr
Brian Herron (07801866678) or Dr
Estelle Healy (07779255420)
Paediatric Pathology – specify either Dr
Caroline Gannon or Dr Daniel Hurrell
P a g e | 225
via RGH switchboard
Cervical Cytology (BCH)
9am – 5pm
Not available
Diagnostic Cytopathology
(BCH & RGH)
9am – 5pm
Contact duty consultant cytopathologist
via BCH or the RGH switchboard
Specialist Services
Immunocytochemistry
(Immunocytochemistry,
8.30am – 5.00pm
Immunopathology,
Electronmicroscopy &
Molecular Pathology)
BCH & RGH
Immunopathology only – contact duty
consultant pathologists (Dr
O’Rourke/Dr Gray/Dr Walsh) via RGH
switchboard or the Immunopathology
Laboratory mobile
Immunopathology
8.45am – 5.00pm RGH
Electronmicroscopy
(Monday-Friday) RGH
Molecular Pathology
(Monday-Friday) BCH
Post-Mortem Service
Mortuary
Mortuary
9am – 5pm
Saturday 9am – 12.30pm
Sunday & Bank Holidays by prior
arrangement only
Laboratory
8.45am – 5.00pm
Adult and Paediatric post-mortems can
be arranged by asking for the covering
Adult or Paediatric Pathologist via RGH
switchboard
Laboratory
as required by Mortuary
P a g e | 226
REQUEST FORM & SAMPLES
Essential & Desirable Criteria
REQUEST FORMS
Essential criteria:
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
Patients full name (first name and surname) * / **
Date of birth
Gender
Health & Care Number and / or Hospital Number
Date
Test request
Patient location (for inpatient hospital & ward required) & destination for report
(Consultant / GP: name; address; telephone number and cypher number if
applicable)
Requesting practitioner details (name & contact details)
Source (ward/clinic/theatre)
Signed by the requesting clinician
Private patient YES/NO
H&C Number (Cervical Cytology)
*or proper coded identifier
**initials are adequate for anonymised samples
Desirable criteria:
1.
2.
3.
4.
5.
6.
7.
8.
Clinical information
Patient’s address including postcode & telephone number
Time of specimen
Request
Clinical comment
Previous histology/cytology report numbers
Clinician’s contact number in the case of urgent or frozen section requests
H&C Number (Diagnostic Cytopathology)
On occasion some of the desirable data may be essential. It is preferable to supply all of the
information detailed to ensure that the patient is not inconvenienced or put at unnecessary
risk due to delay in provision of results.
P a g e | 227
SAMPLE LABELLING
Essential criteria:
6.
7.
8.
9.
Patients full name (first name and surname) * / **
Date of birth
Hospital number and / or H&C Number
Date
Please see below for additional requirements.
Desirable criteria:
1. Nature of sample
2. Time of specimen
3. H&C Number (Diagnostic Cytopathology)
Please see below for additional requirements.
14-3-3 Analysis:
See Appendix 1.
Consent:
Products of conception specimens must have examination consent forms
appropriately completed or they will be returned to source.
KEY POINTS TO REMEMBER
 Supply the correct information in a legible form as this is essential to
ensure that the right result is sent out on the right patient
 A properly completed request form must accompany each specimen
 Hospital or GP practice printed labels are preferred for both request
forms and samples
 Handwritten requests must be complete & legible
 Specimens may not be accepted for analysis in some laboratory
disciplines where the essential criterion in the Minimum Data Set is
not met on either the form or the specimen container
 Sample labelling / request form details must match
P a g e | 228
CRITERIA FOR REJECTION OF SPECIMENS
Cervical Cytology
Specimens may be rejected immediately if:
 Form Received with no specimen
 Discrepancy between the patient details on the form and the
specimen
 No patient details on either the request form or the specimen
 Specimens which have leaked and are insufficient for testing
 Specimens deemed unsuitable for testing by BMS at the point of
testing
 Specimens deemed inappropriate for testing by BMS at the point of
testing
 Specimens accompanied by a request form with insufficient
information to send out a report or to determine which test is required
 Vials which are out-of-date
Molecular Pathology
 Samples will be rejected for analysis upon receipt if the slides are
broken and insufficient material remains for analysis.
 Attempts will be made to complete documentation but, if necessary,
the slides will be returned to the requesting laboratory.


ORDERCOMMS
Electronic requesting and reporting (Ordercomms) is being introduced
across BHSCT. This allows you to select tests on screen and print barcoded order information for attachment to the specimen. In the lab the
bar code can be used to identify the patient and the requested tests. We
hope this will greatly simplify the process of utilising laboratory services.
Please contact the laboratory for further details.
REQUEST FORM DESCRIPTION
Tissue
Pathology
Each biopsy must be accompanied by the Histopathology, Cytopathology
or Immunopathology request form signed by the submitting doctor
(separate Neuropathology request forms are no longer available). Advice
on the fixing and handling of different specimen types can be obtained
from the duty consultant pathologist.
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GUIDELINES FOR PROPER SPECIMEN COLLECTION & FACTORS
WHICH MAY AFFECT THE QUALITY OF RESULTS










Only laboratory approved, CE marked, in vitro devices IVDs, must
be used as primary specimen containers, no substitutes or
improvised containers.
Each sample should be placed in a separate sealed specimen bag.
Collect an adequate amount of specimen. Inadequate amounts of
specimen may yield false-negative results.
All container tops must be firmly & properly closed, leakage
adversely affects not only that specimen but other specimens
sharing the transit.
Specimens must be kept in a cool room awaiting dispatch, not in
the sunlight or near a radiator.
Transit to the laboratory should be prompt and specimens must not
be left in uncontrolled vehicles (hot/cold) for any prolonged period.
If processing is delayed please contact laboratory for sample
storage guidance.
Samples which are of inadequate size, in incorrect tubes, clotted,
or badly delayed in transit may not be processed.
If a sample is unsuitable for testing a report will be sent to the
requestor giving the reason and requesting another sample.
Samples must be packaged and sent so as to comply with the
current regulations on the transport and postage of biological
materials – see Transport Section below.
HIGH RISK SAMPLES
For suspected or known Category 3 pathogens, hazard warning Category 3
pathogen labels should be affixed both to the container and the accompanying
request form. If there is any doubt as to whether a specimen is high risk,
please contact the appropriate laboratory.
Hazard Group 3 is defined as a biological agent that may cause severe
human disease and presents a serious hazard to employees; it may present a
risk of spreading to the community, but there is usually effective prophylaxis or
treatment available.
NB: Hazard warning Category 3 pathogen labels should be affixed to ALL
samples taken from patients with pyrexia of unknown origin (PUO) following
foreign travel.
P a g e | 230
Examples of Category 3 Pathogens
Bacteria:
 Bacillus anthracis (Anthrax)
 Brucella species
 Escherichia coli, verocytotoxigenic strains (e.g: 0157:
H7 and others)
 Mycobacterium tuberculosis
 Mycobacteria other than
tuberculosis (MOTT)
 Salmonella typhi
 Salmonella paratyphi
 Shigella dysenteriae (Type 1)
Fungi:




Blastomyces dermatitidis
Coccidioides immitis
Histoplasma species
Paracoccidioides
brasiliensis
 Penicillium marneffei
Viruses:
 All viral hepatitis (except Hepatitis A)
 HIV
 Sudden Acute Respiratory
Syndrome (SARS)
 Prion Proteins:
 Transmissible spongiform
encephalopathies (TSE) e.g: the
agents of Creutzfeldt-Jacob disease
(CJD): variant Creutzfeldt-Jacob
disease (vCJD)
 Fatal familial insomnia
 Gerstman-Straussler-Scheinker
syndrome
 Kuru
Hazard Group 4 is defined as a biological agent that will cause severe human
disease and is a serious hazard to employees; it is likely to spread to the
community, and there is usually no effective prophylaxis or treatment
available. Please contact the Medical Microbiology team immediately if
you suspect a group 4 pathogen, e.g. Lassa, Marburg, Ebola and CongoCrimean. Under NO circumstances should any samples be taken from
such patients without prior consultation.
TRANSPORT
Details may be found in the Laboratories site on the Belfast Trust
Intranet under ‘Transport of Clinical Specimens to Labs’ (by road).
MINIMUM RE-TESTING INTERVALS
Minimum re-testing intervals do not apply in most of surgical pathology and
diagnostic cytopathology (including fine needle aspiration cytology). In the
case of cervical cytology a repeat cervical smear should not be taken less
than three months after the previous test.
The opinion offered by the pathologist(s) in the report may suggest repeat
sampling if there be diagnostic uncertainty or a requirement for more tissue for
other
more
specialised
tests
such
as
electron
microscopy,
immunofluorescence, molecular testing or flow cytometry.
If further
information is required in this context, the pathologist(s) in question should be
contacted directly.
The Royal College of Pathologists is currently considering its guidance in this
regard.
P a g e | 231
REPERTOIRE
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Surgical Pathology (BCH
& RGH)
General Histo-
Surgical
Biopsies
All specimens for routine investigations must be fixed in 10%
formalin. Specimens received from within BHSCT should be
placed in an adequately sized leak-proof container in
accordance with UN 3373 (supplied by laboratory). For large
specimens consultation with the laboratory is essential to
establish the use of the correct containers which can be
supplied. Sharps/Burn bins must not be used and specimens
received in such will be returned and the incident logged on
DATIX.
The majority of all small diagnostic
specimens are reported between 2-6 days of
receipt (excluding Saturdays and Sundays).
A further 2-4 days is usually required for the
reporting of larger specimens. When special
or further investigations are required an
additional 24-48 hrs is often required.
pathology
Specimens received from other sources must be transported in
SARSTEDT UN 3373 containers which can be purchased
through their website.
There is provision for the electronic reporting
of results to the requesting clinician. This
provision is only granted after a request has
been made in writing to the Clinical Lead.
Every effort is made to ensure that
specimens are promptly processed and that
reports are returned to the requesting doctor
as soon as possible. If an urgent report is
required, clinical users are asked to inform
the laboratory of the circumstances.
Perceived response time may be shortened
by the prompt delivery of specimens to the
laboratory. If an urgent response is required,
it is beneficial to provide a bleep number or
reliable contact number on the request form.
P a g e | 232
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Surgical Pathology (BCH
& RGH)
General Histo-
Frozen
Sections
(including
brain frozen
sections)
The laboratory phone numbers are BCH Ext 41245, and RGH Ext 32170.
Frozen section results are
transmitted verbally within 2
hours of specimen receipt in
the laboratory and followed by
a definitive written report within
the normal timescale of
surgical pathology reporting.
pathology
Arrangements for frozen sections required during planned surgery should be
pre-booked with the laboratory. A contact name and phone number should
also be given. Surgeons are asked to arrange their lists so that the tissue
reaches the laboratory preferably between 8.45am and 4.00pm. For any outof-hours frozen sections, arrangements must be made via the duty pathologist.
The tissue must be fresh, and it is the theatre’s responsibility for delivery
arrangements.
On receiving the specimen at source the porter must be informed that it is ‘an
urgent frozen section’ to be carried to the laboratory immediately. The
accompanying request form must have the Essential Information as detailed in
the request form section of this manual as well as the phone number and name
of the requesting consultant. Where tissue samples have to be couriered by
the Transport Department of another hospital, e.g. the MIH and UHD, it is
critical that these arrangements are made in conjunction with the relevant
Transport Department and the Consultant on duty. Transport arrangements
are the responsibility of the requesting hospital. When a taxi is utilised ensure
the specimen is directed to the correct department. Immediate contact with the
laboratory is required for unplanned frozen sections. Frozen section analysis
is unsuitable for primary diagnostic purposes in many aspects of, e.g. breast
disease or lymphoreticular malignancies or where hazard of infection is
suspected. Also, frozen sections give poorer histological definition than
specimens routinely fixed in formalin. Advice on the appropriateness of frozen
section diagnosis may be obtained by contacting the duty consultant.
P a g e | 233
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Surgical Pathology (BCH
& RGH)
Neuro-
Muscle
Biopsy
Contact the laboratory in advance (RGH Ext 32119) to arrange supply of liquid
nitrogen and EM fixative and to ensure that laboratory staff will be available to
receive specimen.
Muscle biopsy-14 days.
pathology
Usually three separate specimens are required from each muscle biopsy:1.
Histology and Enzyme Histochemistry
Specimen size should be approximately 0.3x0.3x0.3 cm. Place the specimen
on a piece of lollipop stick or tongue depressor and wrap in a piece of saline
“dampened” gauze. Place the gauze and specimen in a dry sterilin jar and
immediately place on ice and despatch to the laboratory.
2.
Electron Microscopy
The specimen can be smaller than that for histology. Place the specimen on a
piece of lollipop stick or tongue depressor and immerse in the EM fixative
provided. Immediately dispatch the specimen to the laboratory.
3.
Specialist Investigation
Specimen size should be similar to that of the piece taken for Histology. Wrap
the specimen in the Parafilm supplied and drop into the liquid nitrogen.
Immediately dispatch the specimen to the laboratory.
Note: - Occasionally a skin biopsy may be required to be taken at the same
time as a muscle biopsy. The Neuropathology Laboratory should be informed
of this at the time of the original call as they will supply an extra container of
EM fixative and a container of transport medium for cell culture. The specimen
for cell culture must be despatched to the Department of Medical Genetics with
the appropriate request form. The other specimen for EM can accompany the
muscle specimens where it will be forwarded to the correct laboratory.
In cases of inflammatory
myopathy a provisional report
can normally be provided in 3
working days.
P a g e | 234
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Surgical Pathology (BCH
& RGH)
Neuro-
The optimum specimen is as follows:-
pathology
Peripheral
Nerve
Biopsy
In cases of vasculitis a
provisional report can normally
be provided in 4 working days.
Otherwise 14 days.
Paediatric
Pathology
Frozen
Sections
Surgical Pathology (BCH
& RGH)
1. Two 1 cm pieces stretched on stiff card on saline dampened gauze
(normal tension to avoid contraction).
One piece for electron
microscopy is cut from the above in the laboratory.
2. Send to laboratory as soon as possible.
Arrangements for frozen sections should be pre-booked with the laboratory
and the paediatric pathologists prior to surgery. Further details are as
specified above for General Histopathology frozen sections.
The surgeons contact number should be clearly written on the request form to
ensure a rapid response time.
Surgical Pathology (BCH
& RGH)
Paediatric
Pathology
Paediatric
Tumour
Biopsies
Tumour biopsies (needle biopsies, incisional biopsies or tumour resections)
should be sent immediately to the laboratory fresh and unfixed. The laboratory
should be contacted prior to sending the specimen in order that preparations
can be made.
The biopsy should be placed in a leak-proof plastic sample container with a
screw-cap lid and into a biohazard bag. For needle biopsies, consideration
should be given to placing the tissue onto paper as smaller samples may
adhere to gauze. The tissue should be kept moist with a small amount of
sterile saline. Place a fully completed Histopathology request form into the
side pocket of the biohazard bag in such a manner as to maintain patient
confidentiality and to ensure that it doesn’t become contaminated in the event
of a leakage. The specimen container and the request form should both be
clearly labelled and appropriate patient data provided i.e. HSC number,
hospital number, ward, clinician, date of birth: if possible, use a PID sticker.
The specimen should be transported to the laboratory urgently to minimise
deterioration of unfixed cells in the sample.
Every effort is made to ensure
that specimens are promptly
processed and that reports are
returned to the requesting
doctor as soon as possible. If
an urgent report is required,
clinical users are asked to
inform the laboratory of the
circumstances.
Perceived
response
time
may
be
shortened by the prompt
delivery of specimens to the
laboratory.
If an urgent
response is required, it is
beneficial to provide a bleep
number or reliable contact
number on the request form.
P a g e | 235
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
Depending on the amount of tissue present, the pathologist will decide what
range of investigations are appropriate (tumour banking, touch
preparations/imprints, genetic testing). In order to obtain the fullest information
possible, as much tissue as possible should be obtained and submitted.
Surgical Pathology (BCH
& RGH)
Paediatric
Pathology
Products of
Conception
All pregnancy losses in the first trimester should be examined histologically to
confirm intrauterine pregnancy and exclude a molar pregnancy. Examination
cannot take place without a completed ‘Consent to Histopathological
Examination and Disposal of First Trimester Pregnancy Loss’ form in addition
to a completed General Histopathology request form. If no consent form is
received, the specimen will be returned to the referring unit until one is
completed. Any residual tissues left after processing for histopathological
examination are communally cremated at Roselawn Crematorium, or returned
to the mother if requested.
Surgical Pathology (BCH
& RGH)
Paediatric
Pathology
Placentas
At present (August 2011), the service is limited to examination of placentas
from babies who are born dead, or who die following delivery. Other placentas
can be examined at the request of a consultant obstetrician or neonatologist.
Send placentas in a leak-proof container fixed in a sufficient quantity of
formalin. A completed ‘Request for Histopathological Examination of Placenta’
form should be submitted with the placenta.
TAT
P a g e | 236
SPECIALTY
Cervical Cytology (BCH)
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Cervical
Samples
Before taking the smear, note expiry date on sample collection vial. Do not use
expired vials. Specimens received in out-of-date vials will not be processed.
Ensure the entire plastic seal is removed from the lid of the vial and discarded
before taking the smear.
National guidelines for cervical
screening suggest that a report
should be available to the
woman within a month of a
smear being taken.
The label on the sample vial should record the forename, surname and date of
birth and/or H&C number to allow matching of the vial with the request form in
the laboratory. Addressograph labels are the preferred option, but labelling IN
PENCIL is also acceptable.
Write the patient’s full name and date of birth on the provided ThinPrep LBC
vial in pencil before starting to take the smear; alternatively, attach relevant
pre-printed label. Check details with the patient in person – especially the
spelling of the surname.
The cervix should be visualized and the smear taken by rotating the cervical
brush 5 times in a clockwise direction.
Immediately rinse the collected material vigorously into the vial. Seal the vial
and send to the lab for processing with a request form bearing all patient
details.
Take time to fill in the request form correctly as samples with insufficient details
will be returned. The details are required both for proper identification of the
patient and for proper assessment of the smear.
After collection and labelling, the sample and request form should be placed in
separate sections of the plastic specimen bag provided before laboratory
dispatch.
Cervical
smears
with
abnormalities or from patients
with a positive history may
take a little longer as these
require special consideration
and reporting by medical staff.
P a g e | 237
Clinical advice on cervical smear taking can be obtained by phoning Dr D
McGibben, Lead Cytopathologist at BCH 028 9504 6141 or Dr G O’Hara,
Consultant Histopathologist/Cytopathologist at BCH 028 9504 7466.
If unsure of how to provide specimens for cytological investigation
please contact the appropriate laboratory for advice (see Phone Numbers
below).
Supplies Requests – Cervical Cytology
Requests for ThinPrep LBC vials, brushes and request forms should be
directed to Screenlink,
Phone/voicemail 00353 1 4605270
Fax 00353 1 4605248
E-mail order@screenlink.net
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SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
Diagnostic Cytopathology
(BCH & RGH)
Diagnostic Cytopathology
(BCH & RGH)
A reporting time of 2-3 days from
receipt of specimen is usual. Priority
is given to hospital in patients. If
required, any specimen can be
examined on an urgent or immediate
basis provided sufficient clinical
information is received for proper
specimen assessment and a contact
phone number for the requesting
physician is made available.
Peritoneal/
Pleural/Peri
cardial
Fluid
Diagnostic Cytopathology
(BCH & RGH)
TAT
Urine
Please send no more than one litre of peritoneal/pleural fluid. This
should not be put in fixative. It should not be sent in a plastic drain
bag or a rigid Perspex drain box but in a sterile, wide-mouthed
container. Essential Information as above.
A freshly voided specimen should be submitted. No fixative should be
used. This should be put in a wide-mouthed container.
Prior to collection of the urine sample the patient should be well
hydrated for 1.5 – 2 hours. During the period of hydration the urine
should be discarded. The next voided urine sample should be
collected and submitted for cytological examination. Early morning
samples are of little value as these show marked cellular degeneration.
Please state on the request form if the patient has been catheterised,
has had any form of instrumentation, has stones in the urinary tract or
is on any form of chemotherapy.
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SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
Diagnostic Cytopathology
(BCH & RGH)
Bronchial
Washings
and
Brushings
The bronchial washings should be submitted in a well-labelled
container. No fixative should be used. The bronchial brush
should be placed in the fixative provided by Cytopathology and
shaken vigorously.
Diagnostic Cytopathology
(BCH & RGH)
Sputum
Three deep-cough early morning samples should be submitted.
Diagnostic Cytopathology
(BCH & RGH)
Fine Needle
Aspirates
If the aspirate is being carried out by the clinician/surgeon and
air-dried and wet-fixed slides are being prepared, they should be
clearly labelled as different staining techniques are used.
Radiologists performing image-guided FNAs of deep sites may
arrange for a BMS to be present by phoning BCH 07557529617,
RGH Ext 33019 or 07824564169. If possible, notification of the
approximate time should be given as soon as it is known.
Contact the laboratory again approximately 15 minutes before
assistance is required.
Arrangements can be made for a pathologist to carry out fine
needle aspiration of superficial sites by phoning Dr Lioe at BCH
028 9504 7351, the Specialist Registrars at BCH 028 9504 1255
or the Cytopathology Laboratory at RGH Ext 33019.
Diagnostic Cytopathology
(BCH & RGH)
ERCP
Brushings
Fixative is provided by Cytopathology. Place brush in pot and
shake vigorously.
TAT
P a g e | 240
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
Diagnostic Cytopathology
(RGH)
Neuro-
Cerebro-
pathology
spinal Fluid
(CSF), Cyst
Fluid
or
Aspirate for
Cytological
Examination
(for CSF 143-3 analysis
see
Appendix 1)
CUSA Fluids
SPECIMEN REQUIREMENT
1. The optimum amount of fluid for cytological examination
is 1 ml or more, preferably not bloodstained.
2. Collect into a leak-proof plastic sample container with a
screw-cap lid and place in a biohazard bag. Place a
fully completed Cytopathology request form into the side
pocket of the biohazard bag in such a manner as to
maintain patient confidentiality and to ensure that it
doesn’t become contaminated in the event of a leakage.
When clinically indicated, both the biohazard bag and
the request form should be labelled with a category 3
(green) sticker.
1. Send as soon as possible to the laboratory to minimise
the deterioration of unfixed cells that the sample may
contain. Store refrigerated at 4C if a delay in transport is
anticipated.
2. CUSA washings to be sent from all neurosurgery cases.
3. Collect into a leak-proof plastic sample container and
send as detailed in 2 and 3 above for CSF, etc.
TAT
CSF-48 hours.
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SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
Specialist Services (BCH
& RGH)
Immunocyto-
All
specimens
For details see specimen types above
chemistry
Specialist Services (BCH
& RGH)
Immuno-
If Immunofluorescence is required for diagnosis, the tissue
sample and/or blood sample must be unfixed.
pathology
Specialist Services (BCH
& RGH)
Immuno-
Dermato-
pathology
logy
Specimens
Dermatology specimens requiring Immunofluorescence usually
consist of a skin biopsy/mucosal biopsy/conjunctival
biopsy/blister fluid/muscle biopsy and a clotted blood sample.
The laboratory will supply transport medium in a leak-proof
container and a leak proof container for the clotted blood
sample/blister fluid sample. These are sent in an outer mailing
container which complies with UN 3373. The outer box is
labelled with the delivery address.
Once the biopsy has been performed please arrange transport
to the laboratory.
Specialist Services (BCH
& RGH)
Immunopathology
Red
Cells
Blood
RBCs from patients with Erythropoietic Protoporphyria (EPP)
fluoresce red when exposed to blue or green light. This auto
fluorescence fades very rapidly and any requests for this test
must be dealt with immediately.
Blood needs to have anticoagulant added. Cover the sample
and transport to the laboratory as quickly as possible (please
phone the Immunopathology Laboratory to arrange this).
TAT
P a g e | 242
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
Specialist Services (BCH
& RGH)
Immuno-
Renal
Specimens
Renal
specimens
requiring
diagnostic
pathology
(Immunofluorescence, Histopathology and Electron Microscopy)
are routed through the Immunopathology Laboratory. When a
renal biopsy is planned, please phone the laboratory to arrange
delivery of the special fixatives kit which contains the requisite
jars of fixatives in the appropriate secondary leak-proof
containers in an outer box. This complies with UN 3733. The
outer box is labelled with the delivery address and contains a
request form. Once the biopsy has been performed please
arrange transport to the Immunopathology Laboratory and
phone to advise that the specimens are in transit.
pathology
If a kit is unavailable the tissue specimen may be placed in
saline and transported as quickly as possible to the laboratory
(please phone the Immunopathology Laboratory to advise that a
specimen is in transit).
Specialist Services (BCH
& RGH)
Electron-
All
specimens
Specimens should be placed in EM fixative which can be
requested from the EM laboratory.
Renal
Specimens
Renal specimens (see above for Immunopathology).
microscopy
Blood
Blood needs to have anticoagulant added. Cover the sample
and transport to the laboratory as quickly as possible (please
phone the EM Laboratory to arrange this).
TAT
P a g e | 243
SPECIALTY
SUB
SPECIALTY
SPECIMEN
TYPE
SPECIMEN REQUIREMENT
TAT
Specialist Services (BCH
& RGH)
Molecular
Pathology
**EGFR
Mutation
Analysis
5x 5-µm unstained sections on uncoated slides plus an HE-stained
section OR Tumour block; Copy of pathology report OR original Specimen
Request Form. In cases of sparse tumour material, serial 5x 5-µM
unstained sections on uncoated slides plus HE-stained sections from
sections OR Tumour block. Current algorithms deal with EGFR and ALK
requests at the same time to save on material (see below).
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 6-9 days (4-7 working days).
**ALK
Translocation
Analysis
2x 3 µm plus 2x4 µm unstained sections on TOMO-branded slides.
These can be ordered initially from NI MPL. Copy of pathology report OR
original Specimen Request Form. In cases of sparse tumour material it is
best to follow the current algorithm, requesting EGFR and ALK together.
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 6-9 days (4-7 working days).
RAS
Mutation
Analysis
5x 5-µm unstained sections on uncoated slides OR Tumour block; Copy of
pathology report OR original Specimen Request Form.
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 14 days (10 working days)
BRAF
Mutation
Analysis
5x 5-µm unstained sections on uncoated slides OR Tumour block; Copy of
pathology report OR original Specimen Request Form.
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 6-9 days (4-7 working days).
2x 3-µm unstained sections on coated slides OR Tumour block; Copy of
pathology report OR original Specimen Request Form; Her2 IHC-stained
slide.
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 6-9 days (4-7 working days).
2x 3 µm plus 2x4 µm unstained sections on coated slides OR Tumour
block; Copy of pathology report OR original Specimen Request Form;
Her2 IHC-stained slide.
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 6-9 days (4-7 working days).
(see appendix
3)
DDISH
Her2
Gastric
HER2
for
P a g e | 244
MSI
5x 5-µm unstained sections on uncoated slides plus a HE stained section;
Copy of pathology report.
Lung Cancer only: Our current diagnostic algorithm encompasses EGFR
and ALK requests for lung adenocarcinoma. Where sample material is
limited, we recommend that the FFPE block is carefully trimmed to expose
the surface of the material on the microtome and the sectioning protocol
as shown below, is followed. The HE can provide “levels” and can be sent
to us along with the blank sections for EGFR and ALK testing. Further
information is available upon request.
Section Number
1
2-4
5
6-10
11
12-15
16
Purpose
H&E
IHC
H&E
EGFR
H&E
ALK
H&E
Cytopathology specimens:- as above from cytoblocks OR cytospins fixed
in Polyethylene Glycol (PEG) or formalin. Please indicate the means of
preparation when making the request.
These items should be placed inside an envelope, Marked Urgent
Pathological Specimen and addressed to:Northern Ireland Molecular Pathology Laboratory,
CCRCB, 97 Lisburn Road, Belfast BT9 7BL
A clinical report signed by a trained
molecular diagnostician and clinical
scientist will be posted in Labcentre
within 14 days (10 working days).
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Appendix 1
CSF 14-3-3 Analysis
Before sending a sample to Tissue Pathology for 14-3-3 analysis the patient's clinician should make
direct contact with the National CJD Surveillance Unit (Tel: 0131 537 1980) in Edinburgh where
they can discuss the patient's clinical picture with one of the unit’s specialist staff. They will give
guidance as to whether the sample should be directed to Tissue Pathology for onward dispatch to
Edinburgh or sent directly. If the Tissue Pathology laboratory receives the specimen, contact will be
made with the National CJD Surveillance Unit to arrange for onward transportation.
When requesting CSF samples for 14-3-3 analysis the CJD Surveillance Unit has produced its own
specially formatted request form which was circulated to the Neurology and Neurosurgical Wards, this
must be completed and forwarded to Neuropathology along with the sample. If a copy of this form
isn't available on the ward the clinician may request an emailed copy to be sent to them by either the
CJD Unit or the Tissue Pathology laboratory (02890632119).
CSF samples requiring 14-3-3 analysis must be sent to the Tissue Pathology laboratory as soon as
possible. For this to happen in a timely fashion the laboratory should be contacted Monday to Friday
8.45am to 5.00pm (02890632119) for arrangements to be made for the dedicated laboratory courier
to collect the sample.
Ideally 0.5-1.0ml of CSF fluid should be submitted for testing. Samples for 14-3-3 analysis must not
be blood stained as this will render the sample unsuitable for diagnosis. If cytological examination is
also required a separate sample should be submitted. The sample/s must be placed in a leak proof
sample container and be clearly labelled with the patient's details and a Category 3 sticker, and
placed in a "Danger Risk of Infection" bag which also contains a pouch where the appropriately
completed and Cat 3 labelled Pathology request form and CJD Surveillance Unit request form can be
placed to accompany the sample and where it will be protected from sample leaks.
Copies of 14-3-3 prion testing results will be forwarded directly to both the Consultant and the
laboratory.
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Appendix 2
Specialty – Post-Mortem Service
Sub Specialty – Adult PM Service
When there is a death in the hospital the Doctor may wish to issue a death certificate or request a
post-mortem. Guidance on death certification and cremation certification is available from the
dhsspsni.gov.uk website in a document entitled “Guidance on death, stillbirth and cremation
certification”. This has been circulated to hospital staff.
In the event that the hospital doctor wishes to have a consented post-mortem, there are consent
forms on each ward. These are triplicate forms and all sections must be filled in. One copy of this
form is given to the relatives. One copy is put in the hospital notes and a third copy is for the
Pathologist. It is very useful if a Doctor, who knows the patient well, fills in a clinical summary form.
This can guide the Pathologist as to the major issues involved in any case. The Doctor may wish to
indicate specific questions that need answered by doing the post-mortem.
The hospital notes, the consent form and the clinical summary accompany the body to the mortuary.
The clinical notes should be available to the Pathologist in the mortuary, but should be returned to the
ward within 24 hours of autopsy.
In addition, there will be an indication on the mortuary forms that a post-mortem is to be performed.
This gives the mortuary staff information as to how to proceed with managing the body of the
deceased.
A death may be reported to the Coroner in the following situations: A doctor did not treat the person during their last illness.
 A doctor did not see or treat the person in the 28 days before they died.
 The cause of death was sudden, violent or unnatural, such as an accident or suicide.
 The cause of death was murder.
 The cause of death was industrial disease of the lungs, such as asbestosis.
 The death occurred in other circumstances that may require investigation.
A death in hospital should be reported to the Coroner if:
There was a question of negligence or misadventure about the treatment of the person who
died.
 They died before a provisional diagnosis was made and the doctor was not willing to certify
the cause.
 The patient died as the result of the administration of anaesthetic.
In the instance that a Coroner’s case has to be performed, consent does not have to be taken from
the family.
Information on Coroner’s post-mortems may be obtained from the Coroner’s Service for Northern
Ireland at www.coronersni.gov.uk/questions.
Ward staff should not make arrangements for the eventual release of a body without first
contacting the mortuary staff to ensure that families are not given unnecessary expectations
of return of a body. Further information may be obtained from the mortuary or the Bereavement
Support Office available during opening hours through the RGH Switchboard.
P a g e | 247
Sub Specialty – Paediatric PM Service
1. All paediatric post-mortems are carried out by the Northern Ireland Regional Paediatric
Pathology Service which is based at RGH and the Belfast Trust Mortuary.
2. Paediatric cases may be consented post-mortems or Coroners post mortems.
3. All children, infants and babies over 12 weeks gestation by size, i.e. crown-rump length 6 cm,
must be transferred to the Belfast Trust Mortuary for PM.
4. To arrange a PM clinical staff must contact the Paediatric Pathology Service. Telephone
contact numbers during working hours are:a. Dr Caroline Gannon
02890638342
b. Dr Daniel Hurrell
02890632369
5. Outside normal working hours cases contact the RGH switchboard (02890240503) and ask
for the duty Paediatric Pathologist.
6. Do not give a time for PM/funeral to the family until you have spoken to the Paediatric
Pathologist. The examination will be carried out as soon as possible but timing depends on
the workload in the Mortuary.
7. It is the responsibility of the referring unit to arrange transport to Belfast Trust Mortuary with
the Trust Funeral Director.
8. The baby must be transferred during Belfast Trust Mortuary working hours which are as
follows:- Monday – Friday 9.00am – 5.00pm . If this is not possible the body should be
transferred early the next working day.
9. If from UHD, LVH or MIH, bodies can be admitted outside these hours as the funeral director
is that of BHSCT.
10. Belfast Trust Mortuary telephone contact number: 02890633679.
11. All bodies must be transferred in a suitable casket, coffin or corrugated plastic box. Larger
infants may be wrapped in a sheet or blanket. If no suitable casket is available advice should
be sought from Paediatric Pathology before transfer.
12. The body and casket MUST both be clearly labelled with the name and DOB of the baby.
13. Any personal items accompanying the remains may be placed in the casket or forwarded
along with it.
14. A body transfer form 1B MUST be fully completed in every case to ensure full traceability.
There are three copies: white, blue and green. The white and blue copies must accompany
the body. The green copy is retained by the ward. Further instructions are on the inside
cover of the book of body transfer forms held in the ward.
15. Documents to be sent with the body to the mortuary are:a. Consented case – Fully completed, clearly signed appropriate consent form (baby or
child), white copy only, and a PM request form giving clinical information.
b. Coroner’s case – SUDI proforma/clinical history and hospital notes. Parental
consent is not required for a Coroner’s PM.
16. Hospital cremation – if the baby is to be cremated by the Paediatric Pathology Service the
appropriate forms should be included with PM documentation.
17. Paperwork should be placed in a sealed envelope labelled with the name and DOB of the
baby and given to the funeral director at the time of transfer. It should not be placed in the
container with the body.
18. All sudden unexpected deaths in infancy (SUDI) and in childhood must be reported to the
coroner and investigated thoroughly. Priority must be given to the preservation of forensic
evidence at all times.
19. When the PM has been completed the Belfast Trust Mortuary will contact the funeral director
of the referring hospital or family to arrange the release of the body.
20. For further information/advice please contact the Paediatric Pathology Service at the numbers
above.
Category 3 Cases
Cat 3 cases (communicable diseases) pose a risk to staff handling the remains, including mortuary
staff, pathologists and designated funeral directors.
If a patient is Cat 3, the remains must be clearly labelled as such by using a Cat 3 sticker, both on the
wrappings around the body and on the accompanying documentation.
P a g e | 248
The designated funeral director who is transferring the remains to the Belfast Trust Mortuary should
be made aware of the Cat 3 status of the remains, but for patient confidentiality reasons, the nature of
the infection need not be disclosed.
Turnaround Times
Autopsy reports should be completed within 12 weeks, although complex cases or those with
peripheral laboratory involvement may take considerably longer.
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Appendix 3
Specialty – Specialist Services
Sub Specialty – Molecular Pathology
Descriptions of Tests
RAS and BRAF
Clinical Rational
Several studies have shown the high prognostic value of KRAS and NRAS mutation response to
Cetuximab treatment in metastatic colorectal cancer. Patients with wild-type KRAS and NRAS
(labelled negative in our results section) are more likely to have disease control on cetuximab
therapy. Patients with KRAS mutation (labelled positive in our results section) may not benefit from
this treatment. Note that all samples for RAS testing are first tested for KRAS. NRAS and BRAF
testing (see below) will proceed only if this first result indicates wild type KRAS status. Known
“hotspot” mutations on KRAS and NRAS genes are identified by COBAS QPCR and Sanger
sequencing (Roche/Life Technologies).
EGFR
Clinical Rational
Several studies have shown the high prognostic value of certain EGFR mutations’ response to
Gefitinib and Erlotinib treatment in adenocarcinoma of the lung. Patients with mutations in the EGFR
gene, specifically exons 18, 19 or 21 (labelled positive in our results section) are more likely to have
disease control on Gefitinib or Erlotinib therapy. Patients with mutations in the EGFR gene exon 20
however, may not benefit from such treatments. Patients with no detectable mutations in the EGFR
gene (labelled negative in our results section) may not benefit from such treatments. Known
“hotspot” mutations on the EGFR gene are identified by COBAS QPCR and Sanger sequencing
(Roche/Life Technologies).
ALK
Clinical Rational
Several studies have shown the high prognostic value of translocations involving the ALK gene in
adenocarcinoma of the lung. Patients with translocations involving ALK are more likely to respond to
Tyrosine Kinase Inhibitor therapy. Patients with no detectable translocation involving ALK (labelled
negative in our results section) may not benefit from such treatments. Translocations involving ALK
are identified by Immunohistochemistry and in situ hybridization technologies (Roche/Ventana).
P a g e | 250
BRAF
Clinical Rational
Several studies have shown the high prognostic value of BRAF mutation response to Vemurafenib
treatment in malignant melanoma and to Cetuximab and Panitumamab in colorectal cancer. Patients
with BRAF mutation (labelled positive in our results section) are more likely to have disease control
on Vemurafenib therapy. In addition, the presence of BRAF mutation is consistent with sporadic
colorectal cancer rather than an inherited defect in MMR in HNPCC. Moreover, in papillary thyroid
carcinoma, BRAF mutations are more likely to have disease associated with extrathyroidal invasion,
lymph node metastasis and advanced stage disease. The presence of the V600E BRAF mutation is
used in the differential diagnosis to exclude Hairy Cell Leukaemia from Hairy Cell variant. . Mutations
on the BRAF gene are identified by COBAS QPCR and Sanger sequencing (Roche/Life
Technologies).
DDISH Her2
Clinical Rational
Several studies have shown the high prognostic value of Her2 overexpression and gene amplification
response to Trastuzumab treatment in breast cancer. Patients with overexpression and gene
amplification of Her2 (labelled positive in our results section) are more likely to have disease control
on Trastuzumab therapy. Patients without Her2 overexpression and gene amplification (labelled
negative in our results section) may not benefit from this treatment. In addition, the DDISH method
detects amplification of the Her2 gene and is used as an adjunct procedure to existing clinical and
pathologic information for prognostic purposes in breast cancer patients and to show that patients are
more likely to have disease control with stage II, node positive breast cancer treated with adjuvant
cyclophosphamide, doxirubicon and 5-fluorouracil (CAF). Amplification of the HER2 gene is identified
using in situ hybridization technology (Roche/Ventana).
HER2 (Gastric)
Clinical Rational
Studies have shown the high prognostic value of Her2 overexpression and gene amplification
response to Trastuzumab treatment in gastric cancer. Patients with overexpression and gene
amplification of Her2 (labelled positive in our results section) are more likely to have disease control
on Trastuzumab therapy. Patients without Her2 overexpression and gene amplification (labelled
negative in our results section) may not benefit from this treatment.
MSI (Colorectal Cancer)
Clinical Rational
MMR-deficient (dMMR or microsatellite instability-high, MSI-H) tumours are found in approximately
15% of all colorectal cancers. dMMR/MSI-H tumours occur in patients with Lynch syndrome, usually
in patients under 50 years of age, and recent Royal College of Pathologist guidelines suggest that all
tumours in patients in this category should be tested by at least MMR IHC along with MSI and BRAF
P a g e | 251
mutational testing. There is also an increased risk for cancer of the endometrium, small intestine,
ovary and other organs. As such, dMMR/MSI testing can be applied to other tumour samples.
If you need further clarification, please do not hesitate to contact:Prof. Manuel Salto-Tellez (m.salto-tellez@qub.ac.uk) Tel. 028 90972718
Dr Jacqueline James (j.james@qub.ac.uk) Tel. 028 90975781
Dr Perry Maxwell (p.maxwell@qub.ac.uk) Tel. 028 90972708
Dr Stephen McQuaid (s.mcquaid@qub.ac.uk) Tel. 028 90972706
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Appendix 4
Referrals to Laboratories outside the BHSCT
Please liaise with the Consultant Histopathologist, Cytopathologist or Neuropathologist as
appropriate.
The CPA accreditation status of referral laboratories is annually checked on the appropriate
laboratory accreditation website by the Tissue Pathology Archivist. Although the possession of
laboratory accreditation is regarded as indicating participation in the histological and
immunocytochemical external EQA schemes, all referral centres are written to on an annual basis
asking for:1. the technical and diagnostic EQA schemes they participate in,
2. confirmation that they maintain a satisfactory EQA standard for continued practice with details
of any poor performance and
3. evidence of their turn-around-times.
Human Papilloma Virus (HPV) Testing in Cervical Cytology
The aetiological role of Human Papilloma Virus (HPV) in the development of cervical cancer is well
established. The NHSCSP has therefore introduced additional High Risk (HR) HPV testing on
selected cervical cytology samples which has the benefit of fast tracking women at risk for treatment
and also reduces unnecessary repeat tests. Samples for HPV testing are dispatched to Altnagelvin
Area Hospital.
HPV Triage
The HPV Triage test is performed on routine cervical samples showing a low grade cytological
abnormality. This includes borderline changes and mild dyskaryosis. A negative (not detected) HPV
test allows a woman to return to normal recall whereas a positive (detected) HPV test initiates a
referral to colposcopy.
HPV Test of Cure
The HPV Test of Cure (ToC) is performed on samples taken following large loop excision of the
transformation zone (LLETZ) treatment for an abnormality, as per request from colposcopy clinician.
Uncertainty of Measurement
In cervical cytology the examination of the sample is used to indicate the presence or absence of
disease. Where biological variation in samples occurs, this can lead to difficulties in interpreting
results. Where there is genuine doubt as to whether cells changes are abnormal an equivocal report
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“borderline changes‟ is issued. This report will initiate either an HPV test on the original sample to aid
patient management or a repeat test.
Within the NHSCSP the sensitivity and specificity of the test is well understood and documented. All
laboratories and screening individuals are monitored and expected to achieve sensitivities in excess
of 90% for all abnormalities and more than 95 % for high grade abnormalities.
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Referral Laboratory Addresses
Department of Cytopathology,
Altnagelvin Area Hospital,
Glenshane Road,
Londonderry, BT47 6SB
Institute of Liver Studies
Kings College Hospital
Denmark Hill
London, SE5 9RS
Department of Pathology,
Basingstoke & N. Hampshire NHS
Foundation Trust
Aldermaston Road
Basingstoke
Hampshire, RG24 9NA
Department of Pathology
Manchester Royal Infirmary
Oxford Road
Manchester, M13 9WL
NCG Pseudomyxoma Peritonei Centre
Basingstoke & N. Hampshire NHS
Foundation Trust
Aldermaston Road
Basingstoke
Hampshire, RG24 9NA
Department of Medical Genetics
‘A Floor’
Tower Block
Belfast City Hospital
Lisburn Road
Belfast, BT9 7AB
Department of Haematology
‘C Floor’
Tower Block
Belfast City Hospital
Lisburn Road
Belfast, BT9 7AB
Department of Pathology
Birmingham Children's Hospital
Steelhouse Lane
Birmingham, B4 6NH
Department of Histology
Level 9
Bristol Royal Infirmary
Malborough st
Bristol, BS2 8HW
Department of Pathology
Cheltenham General Hospital
Sandford Road, Cheltenham
Gloucestershire, GL53 7AN
Department of Pathology
Christie NHS Foundation Trust
Wilmslow Road
Manchester, M20 4BX
Department of Pathology
Queen Elizabeth Hospital
Mindelsohn Way
Edgbaston
Birmingham, B15 2WB
Department of Musculoskeletal Pathology
Robert Aitken Institute
The Medical School
Vincent Drive
Edgbaston
Birmingham, B15 2TT
Department of Pathology
Royal Brompton & Harefield NHS Foundation
Trust
Sydney Street
London, SW3 6NP
Department of Paediatric Pathology
4th Floor
Royal Manchester Children's Hospital
Oxford Road
Manchester, M13 9WL
National Amyloidosis Centre
UCL Division of Medicine
Royal Free Hospital
Rowland Hill Street
London, NW3 2PF
Royal Liverpool University Hospital
Prescot Street
Liverpool
Merseyside, L7 8XP
Department of Pathology
Royal Marsden Hospital
Fulham Road
London, SW3 6JJ
Dept of Pathology
Royal National Orthopaedic Hospital
Brockley Hill
Stanmore
Middlesex, HA7 4LP
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Muscle Immunoanalysis Unit
Lower Ground Floor
Dental Hospital
Richardson Road
Newcastle upon Tyne, NE2 4AZ
Cellular Pathology
L3 New Victoria Wing
Royal Victoria Infirmary
Queen Victoria Road,
Newcastle upon Tyne, NE1 4LP
Department of Pathology
Great Ormond Street Hospital for Children
NHS Trust
Great Ormond Street
London, WC1N 3JH
Source Bioscience
Medical Solutions
Nottingham Business Park
Nottingham, NG8 6PX
Institute of Neurology
Queen Square
London, WC1N 3BG
Nuffield Division of Clinical Laboratory
Sciences
Radcliffe Department of Medicine
Level 4, Academic Block
John Radcliffe Hospital
Headley Way
Oxford, OX3 9DU
Department of Cellular Pathology
John Radcliffe Hospital
Level 1
Headley Way, Headington
Oxford, OX3 9DU
Neurosciences Academic Building
Kings College Hospital
Denmark Hill
London, SE5 9RS
Newcastle Mitochondrial NCG Diagnostic
Laboratory
Department. of Neurology
4th Floor Cookson Building
The Medical School
Newcastle University
Framlington Place
Newcastle upon Tyne, NE2 4HH
LOC: Leaders in Oncology Care
95 Harley Street
London, W1G 6AF
Histopathology Department
Pathology & Pharmacy Building,
Barts Health NHS Trust, 80 Newark Street
Whitechapel, London, E1 2ES
UCL Advanced Diagnostics
1st Floor, Rockefeller Building
21 University Street
London, WC1E 6JJ
Haematological Malignancy Diagnostic
Service
St. James' Institute of Oncology
Level 3 Bexley Wing
Beckett Street
Leeds, LS9 7TF
Department of Pathology
St James’ Hospital
Beckett Street
Leeds
West Yorkshire, LS9 7TF
Department of Cellular Pathology
Neurosciences Centre
Salford Royal Hospitals NHS Foundation
Trust
Stott Lane
Salford, M6 8HD
Department of Cellular Pathology
St George’s Hospital
Blackshaw Road, Tooting
London, SW17 0QT
Department of Cellular & Molecular
Pathology
Antrim Area Hospital
45 Bush Rd, Antrim, BT41 2RL
Histopathology Department, Box 235
Addenbrooke's Hospital
Hills Road, Cambridge, CB2 0QQ
Department Of Pathology Alexander Donald
Building
Western General Hospital
Crewe Rd, Edinburgh, EH4 2XU
Skin Tumour Unit, St John's Dermatology
St Thomas's Hospital
Westminster Bridge Road
London, SE1 7EH
West Midlands Regional Genetics
Laboratory
Birmingham Women's Hospital
Birmingham, B15 2TG
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Regional Genetics Laboratories’
GENERAL INFORMATION
Services
The Regional Genetics Laboratories provide:


Laboratory services for the diagnosis of constitutional chromosome
abnormalities and those acquired in association with cancer, specifically
leukaemias.
Laboratory services for the molecular diagnosis of genetic disorders and for
the identification of carriers of these disorders.
Hours of Service
9:00 a.m. to 5:00 p.m.
Monday to Friday, excluding public holidays
No out of hours service is available
Contact Details/Telephone Numbers
Northern Ireland Regional Genetics Laboratories
Belfast Trust Laboratories
Phone 028 950 48040
A Floor
028 950 48281
Belfast City Hospital
Lisburn Road
BELFAST BT9 7AB
GeneticsLabs@belfasttrust.hscni.net
FAX
028 9023 6911
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Contact Details/Telephone Numbers
Head of Laboratory/Head of Cytogenetics
Mr M Humphreys, Consultant Clinical Scientist
028 950 47915
Mervyn.Humphreys@belfasttrust.hscni.net
Head of Molecular Genetics
Dr Shirley Heggarty, Principal Clinical Scientist
028 950 48281
Shirley.Heggarty@belfasttrust.hscni.net
Genetics Laboratory Manager
Dr Kerry Sweet
02895043386
Kerry.Sweet@belfasttrust.hscni.net
Quality Lead
Dr Claire Byrne
028 950 48155
Claire.Byrne@belfasttrust.hscni.net
Laboratory Secretaries
028 950 48040
028 950 48281
028 950 47844
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Contact Details/Telephone Numbers
CYTOGENETICS
Head of Cytogenetics – Mr Mervyn Humphreys
028 950 47915
Postnatal Cytogenetics enquiries
028 950 40883
Prenatal/Solid Tissue Cytogenetics enquiries
028 950 48126
Cancer Cytogenetics enquiries
028 950 47984
Scientific Staff
Mrs A Logan [Cancer Cytogenetics]
028 950 47984
Dr Geoff Smith [Constitutional Cytogenetics]
028 950 48126
Operational Manager
Miss Judith Briggs
028 950 46113
MOLECULAR GENETICS
Head of Molecular Genetics –Dr Shirley Heggarty
Molecular Genetics Laboratory
028 950 48281
028 950 40878
Scientific Staff
Mrs Diane Beattie [Molecular Genetics, main lab]
Mr P Logan [Cancer Molecular Genetics]
Dr P Hart [Lipid and cardiac Genetics]
Operational Manager
Mr Borghert Jan Borghmans
028 950 47937
028 950 48077
028 950 48301
028 950 48048
Advice
For advice on sending samples and/or appropriate tests please ring the laboratory
secretaries or the individual laboratories as shown above.
The laboratory is happy to assist in ensuring that the most appropriate tests are
carried out (particularly in urgent cases), and to assist with the interpretation of
patients’ test results.
Interpretative comments are added to reports where
appropriate. Clinical referrals are also welcomed.
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Clinical Genetics
The Regional Genetics Laboratories work closely with the Regional Clinical Genetics
Service and this service is also based at A Floor, BCH. Clinical referral is often
required prior to, or following the results of, laboratory diagnosis.
Contact Details/Telephone Numbers
Genetic Nurses/Counsellors
028 950 48010
Consultant Clinical Geneticists
Dr Fiona J Stewart
028 950 48128
Prof Patrick J Morrison
028 950 48177
Dr Alex C Magee
028 950 48042
Dr Shane McKee
028 950 48326
Dr Vivienne McConnell (Clinical Lead)
028 950 48022
Dr Tabib Dabir
028 950 48261
Dr Deirdre Donnelly
028 950 48169
Request Forms
Regional Genetics Laboratory “Medical Genetics” request forms, with integral plastic
sealable envelopes, are available from the department, either singly or in books of
25. These forms have space for patient details, the investigation(s) required, and a
Consent Form. Information about samples types and amounts required is included
on the reverse of the form
A properly completed request form must accompany each specimen.
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Sample Details
The following details must be provided for all specimens:
 The clinical indication for the test.:
o for cytogenetic tests – the reason why chromosome analysis is
requested, including any specific molecular cytogenetic tests required
o for molecular tests - the disease to be tested for and the exact test, if
known
 Patient details
o Name
o Address
o Date of birth
o HCN number
o Hospital / Ward / Patient No.
o Referring consultant / other doctor to receive report copy.
o GP
o Ethnic origin, required in the estimation of recurrence risks
 Family history, (if relevant), including names or reference numbers of related
cases
Patient ID labels should be used, if available
Consent
Current practice is for clinical staff to seek written informed consent prior to genetic
testing or storage of genetic material (cells or DNA). It should be noted that it is the
laboratory’s policy to permanently store excess DNA after the requested tests have
been completed. This provides a resource for future tests which may be required.
Samples may not be processed without appropriate written informed consent.
Please note that all genetic tests requested as part of a private consultation will incur
a charge
Collection and Transport of Samples
 Please use aseptic technique when taking all specimens.
 Please label tubes with
o patient’s full name and date of birth
o date and time of sample
 Details must be complete, otherwise the sample will not be processed and a
repeat will be requested.
Unlabelled samples cannot be processed
P a g e | 261
 Each sample should be placed in a separate sealed specimen bag.
 All samples must be received within 24 hours. If transport is unavoidably delayed
store at 4oC (please record this on the request form).
 Samples must be packaged and sent so as to comply with the current regulations
on the transport and postage of biological materials
 Samples which are of inadequate size, in incorrect tubes, clotted, or badly
delayed in transit may not be processed.
 The following documents are available from the laboratory on request:
o Health & Safety Rules for Porters & Couriers [MF 800 010]
o Pneumatic Tube Transport of Clinical Specimens [LI 800 025]
o Transport of Specimens to the Laboratory [LI 800 026]
High Risk Samples
The laboratory must be informed of any known or potential hazards associated with
samples sent to us



Samples and packaging must be clearly labelled as “biohazard” and the nature of
the hazard details e.g. Hepatitis B, HIV, drug user etc
For some types of sample, and specific categories of hazard, a restricted range
of services may be offered
If in doubt about the appropriateness of material for analysis please contact the
laboratory for advice
Quality Assurance
The Regional Genetics Laboratories are accredited with CPA (UK), CPA No 1973.
Standards for best practice are defined in the Professional and Best Practice
Guidelines produced by the Association for Clinical Genetic Science (ACGS). The
laboratories participate in all available UKNEQAS schemes for the tests offered.
Comments
We welcome feedback on ways to improve our service. Comments from health care
professionals and patients are always welcome and can be sent to the Quality Lead,
Dr
Claire
Byrne,
at
the
above
address
or
by
e-mail
to
Claire.Byrne@belfasttrust.hscni.net
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CYTOGENETICS (Chromosome analysis)
General Information
As chromosome analysis requires tissue culture all samples must be taken under
sterile conditions and transported to the laboratory immediately. Samples taken
outside of normal laboratory working hours should be stored at 4°C (refrigerated),
and sent to the laboratory as soon as is practical.
Samples that are in the wrong container cannot be processed. Tubes are available
from the laboratory.
Appropriate tests, which are clinically necessary, but are not at present carried out
in-house can be arranged as sendaways.
Please contact the laboratory to confirm availability of the test.
Reporting



Reports will be posted out to the referring consultant.
Non-urgent results will not routinely be issued by phone.
Urgent results will be phoned through to the referring clinician only on request.
Additional/supplementary tests
 Cytogenetic cell suspensions are stored during the course of an investigation and
are therefore available for supplementary testing during this time. Cell
suspensions from diagnostic samples received from patients with haematological
malignancies may be kept long term.
 If an additional test is requested after the final report has been issued a new
sample may be required, where possible.
 In cases where an abnormality is detected it is often necessary to test parental
samples before issuing a final report
 In cases where mosaicism is detected a repeat sample or a sample of a different
tissue/site may be requested for confirmation
 All requests for additional tests on existing samples must be made in writing or by
email to the Genetics Laboratory, telephone requests are not accepted until
written confirmation is received.
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Constitutional Cytogenetics
Postnatal enquiries
Prenatal enquiries
028 950 40883
028 950 48126
The Postnatal Section performs cytogenetic analysis for a wide range of referrals,
including infants with multiple congenital abnormalities or dysmorphism; children with
developmental delay or learning difficulties; adults with primary infertility, recurrent
miscarriages (3 or more),or mental retardation; and cases where there is a family
history of chromosome abnormality.
Array CGH: Array comparative genome hybridization (array CGH) is now the firstline test for detecting gene dosage abnormalities in children with developmental
delay (motor or growth), autism spectrum disorder, moderate to severe learning
difficulties, with or without dysmorphic features or congenital abnormalities. This test
has replaced karyotyping and microdeletion MLPA analysis in this referral group.
Samples from infants (<1 year) will be treated as urgent. Non-urgent array requests
however, may for the time being be placed on a waiting list
Recurrent Miscarriage: In line with the Royal College of Obstetricians and
Gynaecologists Green-Top Guideline No.17 (2011) “The investigation and treatment
of couples with recurrent first-trimester and second trimester miscarriage”, the
preferred testing strategy is by the genetic analysis of products of conception (POC)
using molecular techniques to detect chromosome imbalance in the third or
subsequent consecutive miscarriage. Parental testing is now advised only when an
unbalanced structural chromosomal abnormality is found in the POC. Full POC
samples must be sent to the referring hospital’s pathology laboratory, which will then
send appropriate tissue samples to the Regional Cytogenetics laboratory
Please note:
Standard chromosome analysis may not exclude mosaicism.
Chromosome analysis of prenatal samples may not exclude mosaicism or subtle
chromosome rearrangement.
P a g e | 264
See below for sampling guidelines.
Tissue Sampling in Obstetrics & Gynaecology
Recurrent Miscarriage
Genetic investigation of recurrent miscarriage should be undertaken in a couple’s
third or subsequent consecutive first or second trimester miscarriage. Genetic
analysis is carried out on the products of conception (POC) using molecular
techniques (QF-PCR and MLPA) to detect chromosome imbalance.
Parental testing is now advised only when an unbalanced structural chromosomal
abnormality is found in the POC. (RCOG Green-Top Guideline No.17 (2011) “The
investigation and treatment of couples with recurrent first-trimester and second
trimester miscarriage”). In the absence of products of conception material, where
the clinical indications meet the criteria, chromosome analysis of couples may be
requested on blood samples but this should not be considered routine practice.
The miscarried tissue generally consists of the placenta (afterbirth), decidua (lining of
the womb) and blood clot. In a small number of cases there may be fetal remains.
The tissue should be submitted in its entirety to the pathology laboratory in sterile
saline accompanied by a fully completed genetics request form signed by both the
clinician and patient, including details of the history/number of miscarriages. If a fetus
is identified this should be recorded on the form. The tissues should not be placed
in formalin fixative. The specimen should be submitted to the laboratory on the day
of sampling and if this is not possible it should be stored in a fridge overnight.
The POC specimen should be sent to the appropriate pathology laboratory as
follows:
BHSCT and SEHSCT -> Royal Victoria Hospital
NHSCT –> Antrim Area Hospital
SHSCT – > Craigavon Area Hospital
WHSCT –> Altnagelvin Area Hospital
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POC specimens must not be submitted directly to the genetics laboratory as it is
unable to undertake tissue sampling. Any specimens so received will be returned
immediately to the referring Trust, as above
On receipt in the pathology laboratory the Consultant Pathologist takes a sample of
placental villous tissue (2-4mm x 2-4mm) for genetic testing. This is done under
sterile conditions, using a sterile scalpel and needle. The sample is placed in 5ml of
sterile saline or transport (tissue culture) medium and forwarded immediately to the
medical genetics laboratory. If this is not possible, e.g. at weekends, the sample may
be stored in the fridge and submitted as soon as is practicable. The pathologist
should record the sampling and the pathology laboratory reference number. The
remainder of the POC specimen is then formalin fixed for routine histopathological
examination.
These samples are analysed by QF-PCR for common aneuploidies and by MLPA for
subtelomere rearrangements. The results of the genetic tests will be provided to both
the Obstetrician and the Pathologist.
Intrauterine death, stillbirth and neonatal death:
It may be important to undertake genetic analysis when a baby dies in utero, is
stillborn or dies in the neonatal period. If the baby is for post mortem examination the
paediatric pathologist will submit a sample of skin for genetic testing if required. If
there is to be no post-mortem then the clinicians may request genetic analysis.
IUD, stillbirth and neonatal death tissues should be submitted in their entirety to the
appropriate pathology laboratory in sterile saline accompanied by a fully completed
genetics request form signed by both the clinician and patient.
The tissues should not be placed in formalin fixative.
The specimens should be submitted to the laboratory on the day of sampling and if
this is not possible it should be stored in a fridge overnight.
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The tissue specimens should be sent to the appropriate pathology laboratory as
follows:
BHSCT and SEHSCT -> Royal Victoria Hospital
NHSCT –> Antrim Area Hospital
SHSCT – > Craigavon Area Hospital
WHSCT –> Altnagelvin Area Hospital
These tissue specimens must not be submitted directly to the genetics laboratory as
it is unable to undertake tissue sampling. Any specimens so received will be returned
immediately to the referring Trust, as above
On receipt in the pathology laboratory, the tissue of choice is skin and this should be
full-thickness. It must be taken under sterile conditions to prevent bacterial
contamination; the skin should be cleaned with an alcohol wipe and the skin sample
removed using a sterile scalpel and needle. The tissue is put into 5ml of sterile saline
or transport (tissue culture) medium transport medium and should be submitted as
soon as possible to the genetics laboratory, accompanied by a fully completed
request form signed by both the clinician and the mother. When completing the
request form please note that the baby is the patient, however the mother's details
should also be provided.
These samples are analysed by QF-PCR for common aneuploidies and by MLPA for
subtelomere rearrangements. The results of the genetic tests will be provided to both
the Obstetrician and the Pathologist.
For advice re sampling contact the Genetics lab at 02895048126 or Paediatric
pathology at 02890633857.
For enquiries regarding results contact the postnatal section at 028 950 40883
P a g e | 267
Infertility
All referrals from patients for investigation for infertility (including recurrent
miscarriage) MUST be referred by a Consultant Obstetrician/Gynaecologist or
following discussions with the Clinical Genetics Service.
Chromosome Breakage Studies
Chromosome breakage tests for the rare Chromosome Instability Syndromes are
no longer performed by the laboratory. Further information can be obtained by
contacting the laboratory at 028 950 47984 / 028 950 47915
Prenatal Diagnosis
The Prenatal Section performs chromosome analysis from amniotic fluid and
chorionic villus samples (CVS) in pregnancies at increased risk of a chromosome
abnormality.
All prenatal request forms must include gestation.
Prenatal tests on chorion villus biopsies must be pre-arranged by contacting the
Cytogenetics Laboratory on 028 950 48126.
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Quantity/Tube
Sample/Test
Comment
Postnatal
Blood
5ml
For karyotype and/or FISH analysis
Lithium heparin
Microdeletion MLPA
analysis
4-8 ml EDTA
Array CGH
4-8 ml EDTA
[1-2 ml for babies]
[1-2 ml for babies]
Where abnormalities are detected
by array, follow up lithium heparin
or EDTA Blood samples will be
requested on probands and
parents to confirm results and
inheritance by FISH, array CGH or
another molecular method as
appropriate
Prenatal
Amniotic fluid
10-15 ml
For karyotype and/or FISH analysis
Sterile plastic universal
Chorion villus
sample
In transport medium
(supplied by the
laboratory)
By arrangement only
2-4mm x 2-4mm full
thickness skin
For storage and additional
biochemical tests etc as
appropriate
For karyotype and/or FISH analysis
Tissues
Tissue biopsy
Place in sterile
saline/transport medium in
a suitable sterile container
P a g e | 269
Sample/Test
Products of
Conception (POC)
Quantity/Tube
POCs should be sent
direct to associated
pathology lab
Consultant Pathologist
to take appropriate
sample as below and
forward to Genetics
Comment
For QF-PCR and Subtelomere
MLPA analysis
Samples should preferably be
received by Cytogenetics
laboratory on the same day POC
samples are collected. If this is not
possible, samples should be
retained in the fridge at 4oC
overnight
2-4mm x 2-4mm biopsy of
placental villous tissue
Place in sterile
saline/transport medium in
a suitable sterile container
Intrauterine Death
(IUD) / STILLBIRTH
IUDs should be sent
direct to associated
pathology lab
Consultant Pathologist
to take appropriate
sample as below and
forward to Genetics
If you require advice re sampling
contact the Genetics lab at
02895048126
For QF-PCR and Subtelomere
MLPA analysis
Samples should preferably be
received by Cytogenetics
laboratory on the same day
IUD/Stillbirth samples are collected.
If this is not possible, samples
should be retained in the fridge at
4oC overnight
2mmx2mm biopsy of skin
(full thickness)
Place in sterile
saline/transport medium in
a suitable sterile container
If you require advice re sampling
contact the Genetics lab at
02895048126
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Reporting
Sample/Test
Reporting time guideline
Comment
Urgent Blood
10 days
neonate/in-patient infant
<1yr old
Routine blood
28 days
FISH confirmation of
trisomy 13, 18 or 21
3 days
Microdeletion MLPA
28 days
Urgent array CGH
14 days
Routine array CGH
28 days
Postnatal
Usually available in 24
hours
neonate/in-patient infant
<1yr old
Prenatal
Amniotic Fluid
14 days
Chorionic Villus sample
14 days
FISH confirmation of
trisomy 13, 18 or 21
3 days
Usually available in 24
hours
Tissues
QFPCR
28 days
Subtelomere MLPA
28 days
Performed following a
normal QFPCR result
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Molecular Cytogenetic Tests in Constitutional Cytogenetics
Where necessary, additional MLPA/Fluorescence In Situ Hybridisation (FISH)
tests may be carried out to further characterise suspected chromosome
rearrangements, for the detection of microdeletion syndromes and for the detection
of cryptic subtelomeric rearrangements.
FISH tests are carried out on the chromosome preparation obtained from blood
cultures, or occasionally directly from blood smears. However microdeletion testing
by MLPA requires an additional 5ml of blood in an EDTA tube.
In cases where a congenital abnormality suggestive of a chromosome anomaly is
detected on ultrasound scan rapid prenatal diagnosis, using an aneuploidy FISH
test, may be offered. Again no separate sample is required as these FISH tests are
performed on the same amniotic fluid sample received for chromosome
investigations.
Tests available
General FISH tests
 Whole chromosome libraries/paints
 Centromere specific probes (all chromosomes)
 Subtelomeric probes
FISH tests for microdeletion syndromes
FISH testing for microdeletion syndromes has been replaced by MLPA (see below).
However in very urgent cases we may still test for individual syndromes by FISH.







Di George/VCFS (22q11.2)
Prader Willi/Angelman syndrome (15q11-13)
Williams syndrome (elastin gene 7q11.23)
Smith Magenis & Miller Dieker syndromes (17p11.2 & 17p13)
Wolf Hirschorn & Cri du Chat syndromes (4p16.3 & 5p15.2)
1p36 microdeletion syndrome
2q37 microdeletion syndrome
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Other locus specific FISH probes
 SRY Y chromosome (Yp11)
 Steroid sulphatase deficiency (STS) (Xp22.3)
 Kallman syndrome (Xp22.3)
 Rapid Down Syndrome test (LSI21)
Amniocyte - aneuploidy FISH screen




Down syndrome
Edwards syndrome
Patau syndrome
Sex chromosome aneuploidy
This list of probes is not exhaustive as other commercial probes can be obtained on
special request or new probes may subsequently become available.
Whilst most FISH tests are completed within 7 days of receipt they are usually part of
a more complete cytogenetic investigation and results are therefore issued with the
final report. Specific results may however be reported by phone where appropriate
QF-PCR analysis
 Molecular analysis using QF-PCR is used on tissue samples for the diagnosis of
triploidy and aneuploidy for chromosomes 13, 15, 16, 18, 21, 22, X and Y.
MLPA analysis
 Molecular analysis using subtelomere MLPA probe set is used on tissue samples
for the diagnosis of aneuploidy and unbalanced subtelomeric rearrangements
following a normal QF-PCR result.
Molecular analysis using MLPA probe set capable of simultaneous testing for
twenty-three well known microdeletion syndromes.
array CGH analysis
 Microarray analysis using array CGH (available only by referral from or following
discussion with Clinical Genetics Service)
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Cancer Cytogenetics
Contact: 028 950 47984
The Cancer Cytogenetics section examines bone marrow and blood samples for
acquired genetic changes associated with haematological malignancies. Detection of
these abnormalities may aid diagnosis and classification; provide prognostic
information; and help monitor disease status following treatment, during remission,
or after bone marrow transplantation.
The cancer cytogenetic laboratory currently offers a routine diagnostic service for
referrals from patients with known or suspected acute leukaemia, chronic myeloid
leukaemias, myelodysplasias, aplastic anaemias, myeloproliferative disorders,
chronic lymphocytic leukaemias and multiple myeloma. Unfortunately the cancer
cytogenetic laboratory is currently unable to offer a full routine diagnostic service for
all chronic lymphoproliferative disorders, lymphomas and solid tumours.
Samples should preferably be received by the laboratory on the day they are taken.
Since samples are processed by the laboratory on the day after receipt they should
not, when possible, be taken on Fridays.
Myeloma Myeloma Cytogenetics
BONE MARROW ASPIRATES RECEIVED FOR INVESTIGATION OF MULTIPLE
MYELOMA REQUIRE PLASMA CELL PURIFICATION. ANY SUCH SAMPLES
RECEIVED IN THE GENETICS LABORATORY AFTER 12PM ON A FRIDAY WILL
NOT BE PROCESSED
Sample/test
Quantity/Tube
Comment
Bone marrow
1ml
as supplied by laboratory
Take into 5ml transport
medium (medium +
heparin)
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Sample/test
Quantity/Tube
Comment
Blood
5ml
Lithium heparin
Blood should only be sent if the
patient’s disease involves a
significantly raised white cell count
and blast cells have been detected
in the peripheral blood
5ml
For prognostic investigation of CLL
Blood
EDTA
Molecular Cytogenetic Tests in Haematological Malignancies
Where necessary additional Fluorescence in situ hybridisation (FISH) tests are
carried out on bone marrow or blood chromosome preparations from leukaemia
patients to confirm abnormalities detected by banded analysis, or to screen for
important abnormalities associated with specific disease subtypes.
No additional sample is required for these FISH tests
A large range of FISH probes are currently utilised for cancer cytogenetic
investigations, depending on what are commercially available.
Reporting
Sample/Test
Reporting time guideline
Comment
Urgent Marrow/Blood
14 days
New acute leukaemia,
New CML
Routine Marrow/Blood
21 days
Other diagnostic samples
Cancer Cytogenetics
Follow Up Samples
FISH Tests
3-4 days (verbal)
With final report as above
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MOLECULAR GENETICS
Contact 028 950 40878 / 028 950 48281
General Information
The following details must be provided for all specimens:
 The disease to be tested for and the exact test, if known.
o Any relevant test results i.e.
o FH - lipid profile
o CF - sweat & IRT results if known
 The status of the patient
o Affected
o Possible affected
o Possible carrier
 Family history of the disease and names of affected relatives.
 Pedigree if available.
Sample/test
Quantity
Tube
Blood
4-8 ml
EDTA ONLY
[1-2 ml for babies]
Buccal cell
scrapings
By arrangement
In some cases these can be used for
PCR tests if it is not possible to obtain a
blood sample
For other tissue types please contact laboratory to enquire if DNA extraction is
possible.
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DNA Tests
The tests currently available are shown in the DNA Test Table.
Samples will be clinically assessed for sendaway tests to another laboratory for other
conditions if the test is approved by the UK Genetic Testing Network www.ukgtn.org.
These tests can be very expensive and the laboratory reserves the right to request
that referrers pay for these tests. Samples will be prioritized based on clinical need
and may be placed on a waiting list. Where specific Testing Criteria are defined by
the UKGTN, referrers MUST ensure that patients meet these criteria and that the
appropriate forms are supplied with the sample.
For some tests samples will only be processed if the person has been seen by the
Clinical Genetics Service. This is the case for:


Predictive tests for Huntington disease
Predictive tests for inherited cancers
For genetic testing for cardiac disorders such as LQT and HCM referrals will only be
accepted from consultant cardiologists who specialise in inherited cardiac disorders
or from clinical geneticists.
Reporting
The estimated reporting time is given as an approximate guide to availability of
results and applies from the date of receipt of the sample.
Reports will be posted out to the referring consultant. Results will NOT routinely be
issued by phone. Urgent results can be phoned through to the referring clinician on
request.
Additional/supplementary tests
 If a previous sample has been submitted with consent for DNA storage
supplementary tests can be requested without submitting a further sample
 It is, however, advisable to check with the laboratory to ensure that sufficient DNA
of good quality remains in storage
 All requests for additional tests on existing DNA samples must be made in writing
or by email to the Genetics Laboratory, telephone requests are not accepted until
written confirmation is received.
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DNA TEST TABLE
Disease
Cystic Fibrosis
Tests available
Time to
report
Comment
4 wks
Detects 90% of mutations
Fluorescent sequencing / MLPA
>6
mths
Full exon screen on patients with a
confirmed diagnosis.
MLPA for all exons
4 wks
Detects all whole exon deletion and
duplication mutations (65 -80% of all
dystrophin mutations
Range of intragenic microsatellite
markers
4 wks
Carrier testing by linkage and
Fluorescent sequencing of
4 wks
Detects the most common p.G307S
and p.I278T mutations
3 mths
MLPA analysis done in batches
approx every 6 mths.
4 wks
Detects normal and intermediate
alleles (<56 repeats) and lower end
of premutation range (<80)
CF-EU2 test for 50 mutations and
the intron 8 poly tractT
c.1210-12(5/7/9T)
Duchenne/Becker
Muscular Dystrophy
Homocystinuria.
Cystathionine
heterozygosity exclusion
exon 8
B synthatase
Fabry disease.
Fluorescent sequencing
-galactosidase A
All exons 1 – 7 + MLPA
Fragile X / FXATS
Fluorescent triplet repeat PCR
Fragile X tests no longer carried out
as a screening test.
Now only performed following a
normal aCGH result, and only in
patients with relevant clinical
features, or family history as
appropriate
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DNA TEST TABLE
Disease
Tests available
FMR1 AmplideXTM kit
Time to
report
2 mths
Comment
Detects normal, pre and full mutation
alleles
Fragile X tests no longer carried out
as a screening test.
Now only performed following a
normal aCGH result, and only in
patients with relevant clinical
features, or family history as
appropriate
Friedreich ataxia
6 wks
Detects most affected patients and
carriers
Fluorescent triplet repeat PCR
2 wks
Detects normal alleles (<38 repeats)
and small mutation alleles (50-70
repeats)
TP-PCR
2 wks
Detects larger expansion mutations
but cannot determine the size of
expansions
Long range PCR
2 mths
Detects small expansion mutations
Fluorescent triplet repeat PCR
2 mths
Dominant ataxia screen
Triplet repeat PCR Short range, TP-PCR and Long
range PCR assays
Myotonic dystrophy
SCA 1,2,3,6,7,17
SCA7 TP-PCR
DRPLA
Fluorescent triplet repeat PCR
2 mths
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DNA TEST TABLE
Disease
Huntington disease
Tests available
Fluorescent triplet repeat PCR
TP-PCR
Time to
report
2 wks
from
second
sample
Comment
Detects normal alleles (<36 repeats)
and mutation alleles (>35).
Predictive tests require two separate
samples [accepted from clinical
genetics only].
Diagnostic tests carried out on a
single sample
Kennedy syndrome
Fluorescent triplet repeat PCR
4 wks
Detects normal alleles and mutation
alleles
Prader Willi /
Angelman
syndromes
MS-MLPA dosage and methylation
test
6 wks
Detects microdeletions and
heterodisomy and imprinting
mutations.
Fluorescent microsatellite
multiplex analysis
Confirms mechanism in affected
cases – requires parental samples
HMSN / CMT1A /
HNPP
MLPA
2 mths
Detects common PMP22 deletions
(HNPP) and duplications (CMT1A)
Identity testing /
Maternal
contamination assay
in AFC cultures /
Human identity testing - multi locus
multiplex STR assay
2 wks
Identity testing only performed to
confirm other test results.
Turner syndrome Ytest
Y specific STS multiplex
8 wks
Gonadoblastoma susceptibility critical
region
Fetal sexing
Y specific PCR
5 days
Rapid test on amniotic fluid
SRY, SY54, SY81
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DNA TEST TABLE
Disease
Phenylketonuria
Tests available
Time to
report
Comment
3-6
mths
Detects majority of mutations >95%
2-3
mths
Detects ~99% of N. Irish mutations
Carrier testing for relatives of PKU
patients
2-3
mths
Various single mutation tests
Family mutation testing
4 wks
Predictive testing for known family
mutations
FH Biochip assay – basic FH
screen
2 mths
Detects ~70% of FH causing
mutations within the UK and Ireland
3 mths
Detects >90% in definite families
/25% in possible FH families
Diagnostic mutation screening
Fluorescent sequencing
All exons of PAH gene
Partner carrier testing
Exons 2,3,7,8,9,10,11 & 12
Familial hypercholesterolaemia &
FDB
38 LDLR, 1 ApoB, 1PCSK9
mutations
Fluorescent sequencing All exons of LDLR gene +
promoter, + ex7 PCSK9 + ApoB
binding region of ex 26
+MLPA for LDLR
PCSK9 sequencing
LPL deficiency
Full sequencing of LPL
3 mths
6-12
mths
Severe hypertriglyceridaemia
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DNA TEST TABLE
Disease
Tests available
Time to
report
Familial defective
ApoB
R3500Q mutation
4 wks
ApoE genotyping
E2/E2 only
4 wks
Hereditary
Connective Tissue
Disorders
Next Generation Sequencing of
panel disease genes implicated in
CTDs.
Comment
Type III hyperlipidaemia
-
For current Turnaround times please
contact the laboratory
3 mths
Patient should meet the specified
criteria and have seen cardiologist /
clinical geneticist
ACTA2, FBN1, COL3A1,
MYH11,TGFBR1, TGFBR2,
SMAD3
Long QT syndrome
Fluorescent sequencing screen
KCNH2, KCNQ1, KCNE1, KCNE2
all exons
+ SCN5A on request
MLPA for above genes
3mths
Family mutation test
4 wks
Brugada syndrome
Fluorescent sequence screen
SCN5A all exons
3 mths
HCM
Fluorescent sequence screen
3 mths
MYH7, MYBPC3, TNNT2,TNNI3
Patients need to meet the specified
criteria and have seen cardiologist /
clinical geneticist
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DNA TEST TABLE
Disease
FAP colon cancer
Tests available
Family mutation test
Fluorescent sequencing for APC
exons 15 hot spot
Time to
report
4 wks
4-6 wks
Comment
Predictive testing for known family
mutations [accepted from clinical
genetics only]
Detects common 1309 & 1061
deletion mutations
Family mutation test
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
Mutation screening by fluorescent
sequencing
2 mths
Detects coding region and splice
junction mutations
MEN IIa and FMTC
Family mutation test
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
MEN IIb
Family mutation test
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
BRCA 1 / 2
Mutation screening by fluorescent
sequencing + MLPA
2 mths
Detects coding region and splice
junction mutations
Family mutation testing
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
Family mutation testing
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
MUTYH
Hereditary nonpolyposis colon
cancer
(HNPCC)
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DNA TEST TABLE
Disease
Tests available
MLH1, MSH2,
Time to
report
3 mths
Detects coding region and splice
junction mutations
6 wks
Detection of exon deletions and
duplications
fluorescent sequencing
MPLA analysis MLH1 / MSH2
MSH6 / PMS2 sequencing
PGL1
Comment
3 mths
SDHD Family mutation test
4 wks
Predictive testing for known family
mutations [accepted from clinical
genetics only]
4 wks
Familial expansile
osteolysis (FEO)
RANK84dup18 mutation
DNA storage
Extraction of DNA from whole
1. OATs to other UK labs.
o
blood and storage at <20 C.
2. Rare conditions
CONSENT REQUIRED FOR DNA STORAGE
3. Undefined syndromes
CONSENT REQUIRED FOR DNA STORAGE
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PNEUMATIC TUB TRANSPORT OF CLINICAL SPECIMENS
The main hospitals in the Belfast Trust have pneumatic specimen transport systems
which transport specimens in pods from many of the wards to the local on site laboratory
or to a central dispatch point from where specimens are shipped off site by road to
another laboratory.
There is a cross site pneumatic link between the BCH and RVH but the capacity is low
so it could quickly get back logged and result in significant delays. It’s use, and access
to it, therefore is restricted.
This cross site system is only for urgent specimens, it is usually used by laboratory
reception and by the laboratories themselves to forward urgent specimens between
regular van run times, at night time and at weekends, instead of taxis.
Any specialist unit regularly requiring a rapid transfer of specimens to a laboratory on the
other site can make a special request to the laboratories for access to use this directly
Not all laboratories and destination stations are manned 24/7, Where a station is closed
the pods will be re-directed automatically to a 24/7 station where staff will ensure the
material is forwarded to the intended destination in the next appropriate taxi or van run.
Care must be taken to correctly key in the destination station code, if a pod is sent to a
non laboratory destination by mistake it could take a significant time for this to be noticed
and for the recipient to return the full pod to the original sender.
The Laboratory gives the following Guidance on the operation of the Pneumatic
Air-tube Transport System and on what materials may be sent by this means.
Users need to be aware that the Pods used in the system are not the ones the
manufacturer of the system (Summetzberger) designed for laboratory use. They are in
fact document pods and are not leakproof. Care must be taken therefore to ensure
specimens do not leak or get damaged in transit.
Specimens transported in pods by pneumatic tube are subjected to a lot of shaking and
knocking around if they are not well restrained within the pod.
A lot more knocks than when transported by hand by porters
Pneumatic transportation will likely exaggerate any effect of improper packaging and if
leakage occurs may result in a major and expensive clean up since it is likely to leak out
of the pod into the tube network.
Because of the more severe physical strains of pneumatic transport it is all the more
important that the basic rules of specimen packaging are followed.
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You MUST always
 Use a rigid leakproof primary specimen container which is CE marked and
labeled as an ‘In vitro device’ ‘IVD’ or
Containers specifically issued by the laboratories for specimens. (Do not use
improvised or alternate containers).
 Use the correct container for the specimen type, e.g. faeces should only be sent
in faeces containers, the ones with the integral spoons (currently blue lid)
 Securely tighten lids (see attached sheet)
 Properly seal primary container in the request form bag
 Only pack one specimen per request form bag (except swabs where limited
multiples are permitted, provided the bag can be properly sealed)
 Damaged or cracked, containers must not be used
 Containers must not be more than ¾ full or must not be filled above indicated fill
line where there is one.
 Ensure there is adequate padding in the pneumatic pod (a minimum of two
polystyrene bungs pushed down to restrain load)
On the last point the maker’s design specification is for the use of two tightly fitting
polystyrene bungs, one in each end of the pod, these are pushed in to constrain the
contents from being thrown around during transit and to cushion the contents against
impact with the hard ends of the pod when they accelerate, corner or brake.
CORNER
ACCELERATE
DECELERATE/BRAKE
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Securing the lids of screw top 30 ml Universal Containers before Transporting
A = point at which bottle first touches rubber seal
B = point where lid is securely locked and sealed
URINES & BODY FLUIDS
OR
A
B
LIDS MUST BE TIGHTENED BEYOND
POINT OF FIRST CONTACT WITH LID
SEAL AS INDICATED IN DIAGRAM
0
~30
~1cm
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FAECES
Blue lidded faeces containers have a different
type of sealing mechanism.
The Distance from first bite point to securely
closed is shorter, an angle of 150 or 0.5cm
Specimen Types
Most routine Blood and Microbiology Specimens can be sent in the pneumatic
system, provided the basic instructions above are strictly adhered to.
This includes:Swabs, Urines, Blood Cultures, Sputum, Faeces, blood in Vacuette containers.
The only type of specimen container that cannot be used is glass, a lot of users do
not realize that the blood culture bottles are in fact plastic and can be sent in the
pneumatic system.
Specimen Volumes
100ml glass bottles of body fluids must not be sent in the pneumatic system, but in
most circumstances it is not necessary to send such large volumes.
The Microbiology Consultants have agreed that a maximum of two 20 ml plastic
universal bottles (white top Sterilin type bottles) are sufficient for most situations
including Pleural Fluids, CAPD samples and other body fluids
Body fluids can only be sent in the system when packed in singles or multiples of
20ml in universal bottles. Body fluids must not be sent in the larger 50ml sputum
pots.
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Larger volumes or specimens in glass containers must not be sent in the pneumatic
system.
Damaged Pods
Pods with any damage to the lids, body or outside seals must not be used, they are
likely to stick in the system and will shut down sections resulting in significant delay
of any other specimens in the system at the time and specimen transport will have to
fall back onto much slower portering until the system is fixed. Damaged pods should
be wrapped in a bag and returned to the labs with a note, these will be forwarded to
the control room for repair and will return to the ward in due course using its coded
chip.
Pharmacy Pods
Pneumatic Pods for Pharmacy use are designated by a Green Band at each end
while those for Laboratory use are designated by a Red Band at each end.
Laboratory specimens must never be sent in green pods and pharmacy requests
and supplies must never be sent in red pods.
There is a serious control of infection issue if using the same pods. Laboratory
specimens may contain Health Care Associated Infections such as MRSA or Cdiff.
These could leak unnoticed or otherwise contaminate the pod. If Pharmacy supplies
were then sent in the same pod and were handled and taken into a clinical area on
the presumption of being ‘clean’ these infections could be spread. Pharmacy pods
are longer and usually have no installed padding. Consequently laboratory
specimens in these pods are more frequently damaged or leak in transit.
Restricted and Prohibited Specimens
Users must not send specimens where there is a high element of Risk
There are two types of Risk
1.High Value / Fragile Specimens
Where there is a risk of loss, delay, and/or consequent damage to a high value
specimen that may be difficult to replace or difficult to repeat e.g.
CSF
Biopsy
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The pneumatic system does have outages, pods at times can be snagged inside the
system during an outage. If you require an urgent clinical result for guidance and
could not easily repeat the specimen if the first specimen got lost or stuck in the
system then you should request a porter transfer.
Some Clinical Chemistry tests such as blood gas analysis, which must travel on ice,
are unsuitable for the pneumatic system and the knocks may also interfere with the
results.
2.Hazardous Specimens
Hazardous
Leakages and breakages occur in any transport system, but leakage in pneumatic
systems are much more difficult to clean up and decontaminate.
The pods we use are not leakproof, if a specimen were to leak from a patient with a
Hazard Group 3 (Containment Level 3) organism, the entire tube system could be
contaminated. If noticed this could close down the system for several days to
decontaminate, if unnoticed it could spread infection.
Specimens from Hazard Group 3 (Containment Level 3) patients MUST be double
bagged and MUST be labelled as ‘Category 3 Risk’ with a suitable sticker or
Biohazard label so that prompt action can be taken in the event of a leak being
noticed on receipt.
Examples include patients with TB, HIV, HTLV, Hep B C E G, Brucella, Typhoid,
Dysentry & verotxigenic E.coli O157
There are no exceptions, if not double bagged and not clearly labeled these
specimens are prohibited.
Very Hazardous
Specimens from patients with or suspected to have a Hazard Group 4 (Containment
Level 4) organism are totally prohibited from the pneumatic system. Examples
include, rabies, haemorrhagic fevers, SARS, Avian Influenza, Lassa Fever, Ebola
virus etc.
Even for portering and road transport these materials require special arrangements,
so advice and guidance from the Microbiology/Virology consultants and their
Transport Advisors must be sought first.
Training
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It is strongly emphasized that it is always down to the sender to be trained in the
procedures related to the operation of the pneumatic tube and to keep up to date
with subsequent changes in practice.
They should be familiar with and have read any instructions on the transport station
and should check regularly in case the instructions have been updated.
As for any procedure the sender must decide if they are sufficiently trained,
competent, and up to date with procedures before carrying out the transportation
procedure.
The sender must make a brief mental risk assessment of what they are about to do,
taking account of the guidance above, any update training they have received and
considering the possible consequences, they must then decide on the appropriate
action to take.
The fall-back position, if in doubt, is to consult the receiving laboratory first.
As for any procedure Ward Managers must maintain records of staff training and
refresher training in relation to the pneumatic tube. In house, on the ward, training is
acceptable but must be evidenced.
Training must be supported by clear written instructions kept at ward level and
available to all staff using the system.
Wards must not rely on familiarity with pneumatic systems in other hospitals being
directly transferrable to the Belfast Trust system, particularly for recently recruited
staff, agency staff and trainee/placement staff.
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TRANSPORT OF SPECIMENS TO THE LABORATORY
There is a legal responsibility and a duty of care on anyone who dispatches clinical
material (diagnostic specimens) to the Belfast Trust Laboratories, (by whatever
means, hospital van, courier, taxi, post, internal portering, pneumatic chute)
The legal responsibility is to ensure that the specimens are packaged and labeled in
compliance with the relevant road transport regulations (ADR/CDG). There is a
further legal responsibility under COSHH regulations, since clinical materials may
contain infectious agents, to ensure that the materials do not leak or injure anyone
involved in the transportation or the wider public and environment.
The duty of care ( to the patient) is to ensure that the transport conditions do not
damage the material being sent for testing or otherwise interfere with the validity of
the test results, and to ensure the specimen reaches the laboratory in good condition
within an appropriate time frame for good clinical management of the case.
This requires








Submission of the correct type of specimen, in the correct container for the
test required
correct addressing of the specimen/request to the correct laboratory on the
correct form.
full matching PID details on specimen and form
a clear statement of the test required
Indicate if an unusual or fragile organism is suspected as the causal agent,
these may not be isolated by normal testing protocols and may require special
media, special isolation conditions and prolonged incubation
a clear statement of the nature/site of the specimen
a clear statement of the relevant clinical details and history
details of the patient location (where results are to be sent to)
If any of the above are missing this may result in snagging, which may delay
transport to the correct lab, may cause wrong tests to be performed, may result in
delays in reporting
In addition the following transport related factors may cause delays or impact the
quality of results


All container tops must be firmly and properly closed, leakage adversely
affects not only that specimen but other specimens sharing the transit
Only laboratory approved, CE marked, in vitro devices IVDs, must be used as
primary specimen containers, no substitutes or improvised containers
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





The date and time of collection should be clearly stated (24 hr clock) because
some fragile organisms must reach the lab and be plated within a short
timeframe if there is to be a chance of isolating them. E.g. Neisseria
meningitidis or N. gonnhorea
Specimens must be kept in a cool room awaiting despatch, not in the sunlight
or near a radiator
If specimens cannot be shipped until the next day they should be stored in a
fridge at 2-8C with a max/min monitor to ensure this range is maintained.
Specimens must not touch cooling plates which may frost the specimen
URT specimens must be stored in a cool room 10-16C because some of the
significant URT pathogens will die off at 2-8C
Transit to the laboratory should be prompt and specimens must not be left in
uncontrolled vehicles (hot/cold) for any prolonged period.
The desired situation is that all vehicles used to transport specimens should
have adequate active heating/cooling systems in their goods compartment to
ensure the temperature range is maintained between 8 to 22C and this should
be considered in any future rounds of transport procurement or service
contracts with carriers. In the interim mitigating action should be taken by way
of insulation of specimens in transit with or without cool paks
The laboratories are not responsible for nor do they have any managerial control
over the transportation of specimens between the shipper and ourselves. We can
only give advice on good practice to those who send (ship) to ourselves.
the strong recommendation by Belfast Trust Microbiology is that all patient
Clinical Specimens should be considered as potentially infectious and must
therefore be categorised at the very minimum as UN3373 Biological
Substance Category B and be packed and labelled according to Packing
Instruction P650 in the ADR/CDG regulations.
If fully compliant with P650 then the package, the transport vehicle and the driver are
not subject to further specific requirements under ADR.
THIS EXEMPTION MUST ALWAYS BE USED
If the packaging is not P650 compliant then there is no exemption from the full
ADR/CDG regs, and the shipper and the driver will probably be found in breach of a
number of transport regulations and liable to prosecution.
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Shippers Responsibility for Transport
Any Unit, Hospital, Clinic, GP Practice or Trust transporting specimens by road
(which includes postal services) should take professional advice and guidance on
the packaging and labeling of any materials they hand over for transportation
because they are technically the shipper and are therefore legally responsible for
compliance. Any staff involved in the packaging and shipment of Clinical Specimens
should be appropriately trained for their role and this should be documented.
Specimens transported on public roads are subject the Carriage of Dangerous
Goods and Use of Transportable Pressure Equipment Regulations 2011 (CDG 2011)
and the European agreement (“Accord européen relatif au transport international des
marchandises Dangereuses par Route”, (known as ADR) (ADR 2011) which
together regulate the carriage of dangerous goods by road.
Specimen transport is also subject to further guidance issued by the competent
authority for the UK, the Department for Transport, the key document is ‘Transport of
Infectious Substances (DfT) 2009:
A guidance document produced by the
Department for Transport, the Civil Aviation Authority and the Maritime and
Coastguard Agency’
Individual couriers such as the Post Office may issue their own additional
requirements and restrictions, which must be complied with.
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Overview of Packing Instruction P650
Users MUST consult the full packing instruction, this is only an overview. The
packaging standard comprises 3 layers, two leakproof layers, and a third outer rigid
layer which provides protection against impact.
1. PRIMARY CONTAINER (LAYER #1)
2. REQUEST FORM POUCH
Rigid, leakproof, lid tightened.
Individual, Clear plastic envelope, attached to appropriate colour
In Vitro Diagnostic Device (CE marked)
coded form, sealed
This device does not meet the criteria for a leakproof container. It
is a device to direct the specimen to the correct destination and
keep specimen and request together
.
.
Haematology
Clinical
Chemistry
Microbiology
DESTINATION
LABORATORY
Haematology
Clinical
3. OPAQUE/CLEAR SPECI-BULK BAG (optional)
Where large numbers of specimens are being sent samples are sorted at source and
streamed by colour code into pre addressed and colour coded bags supplied by the
laboratory. Sorting and streaming at source speeds specimens through reception
areas.
For road transport purposes this does not meet the criteria of a leakproof layer of
destination laboratory
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4. SECONDARY LEAKPROOF CONTAINER (LAYER #2)
.
UN33
5. TERTIARY RIGID OUTER (LAYER #3)
Road transport outer box
Each specimen does not have to be individually packaged
in its own separate three layer system
BIOLOGICAL SUBSTANCE, CATEGORY B
Several specimens or multiple specimens can be packed
together in a bulk transport system provided the above
model of the three layer configuration is complied with.
There should be sufficient absorbent material between the leakproof layers to absorb
the leakage of one of the containers.
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There should be sufficient padding to ensure specimens do not rattle around loose
inside the package and that multiple specimens are cushioned against knocking
against each other.
The outer container should be marked with the details of the shipper and consignee
and the following symbol and shipping name should be on the outer layer as well as
two sets of this way up arrows on opposite surfaces. Four faces of box shown
below:-
To
UN3373
From
BIOLOGICAL SUBSTANCE, CATEGORY B
For road transport the packaging systems must be certified by the maker/ issuer to
be compliant with the ADR/CDG standards for road transport of UN3373 and must
be used with all the components, and only those components, tested and stipulated
by the maker / issuer as meeting compliance. Alternately the higher standard UN
type approved packaging for Class 6.2 Dangerous goods can be used. These are
independently tested and marked in the style of
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4GU/CLASS 6.2/06 / GB / 2815
Some trusts use self approved, home designed, bulk transport systems in vehicles,
these are suitable if they are properly risk assessed as such but it must be pointed
out that leakproof means through 3600, and with liquid in contact with the access
point/lid. A common misunderstanding is that a loose lidded container, essentially a
trough or bucket is leakproof, this is not leakproof within the road transport regs.
The primary specimen container must be a CE marked ‘in vitro device’ IVD approved
by the laboratory for the particular test requested, not a substitute nor an alternate
improvised container. Other containers may not be sufficiently leakproof or robust
enough to withstand the rigors of transport and will invalidate the compliance of the
packing system.
Users must be mindful that if the lid of the primary container is not properly secured
then it is not leakproof and the entire packing system is not compliant for road
transport. Users must make sure lids are firmly tightened and not rely on patients
having done so when they collect their own specimens.
The transport packaging system must be adequately secured within the vehicle
against the normal rigors of road transport or minor road traffic accidents. It must not
be transported in the passenger compartment of the vehicle.
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Transport of High Risk UN2814 Infectious substance affecting humans,
Category A
There is an important exception to the above general rule where a specimen is
transported from a patient suspected or known to have a highly infectious disease,
one identified as a Containment Level 4 agent by the Advisory Committee on
Dangerous Pathogens. These high risk materials must not be transported to Belfast
Trust Microbiology or any other Trust Laboratory or premises without first consulting
with Microbiology or Virology (these will be highly infectious viruses). These will
require a specialist courier, they must not be transported by post, hospital van or
taxi.
These highly infectious materials are categorised as
UN2814 Infectious substance affecting humans, Category A, they must be packed,
labelled and documented according to Packing Instruction P620 they will require a
specialist courier and are further subject to the Anti Terrorism Crime and Security Act
(ATCSA 2001) as amended 2007, Which defines them as High Consequence
Dangerous Goods and requiring security plans for transport.
Only Belfast Trust Microbiology/Virology Laboratories have the necessary
arrangements and security plans in place with a trained specialist courier for this kind
of shipment and will make the arrangements for the specimen to be collected and
transported to their site to prepare to forward to a Containment Level 4 facility in UK.
As a general rule if there is sufficient suspicion for a clinician to request testing for a
CL4 organism, (even if only for exclusion, and even if still awaiting test results), for
that material and any other specimens being sent for any laboratory testing to be
considered as potentially UN2814, Category A and these should be shipped
accordingly.
For laboratories shipping cultures of CL3 organisms to the Belfast Trust Laboratories
for further testing, many of these cultures should be shipped as UN2814 but some
have derogations to be shipped instead as UN3373. It is the responsibility of the
shipping laboratory to consult the relevant regulations, DfT guidance, and take
advice from a DGSA before shipping.
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Securing the lids of screw top 30 ml Universal Containers before
Transporting
A = point at which bottle first touches rubber seal
B = point where lid is securely locked and sealed
URINES & BODY FLUIDS
OR
A
B
LIDS MUST BE TIGHTENED BEYOND
POINT OF FIRST CONTACT WITH LID
SEAL AS INDICATED IN DIAGRAM
0
~30
~1cm
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FAECES
Blue lidded faeces containers have a different
type of sealing mechanism.
The Distance from first bite point to securely
closed is shorter, an angle of 150 or 0.5cm