CHHS16/004
Canberra Hospital and Health Services
Clinical Procedure
Blood Culture Collection
Contents
Contents ..................................................................................................................................... 1
Purpose....................................................................................................................................... 2
Alerts .......................................................................................................................................... 2
Scope .......................................................................................................................................... 3
Section 1 – Peripheral blood culture collection ......................................................................... 4
Section 2 – Collection from a Central Venous Catheter (CVC) or Percutaneously Inserted
Central Catheter (PICC) .............................................................................................................. 6
Section 3 – Collection from an Implanted Venous Access Devices (e.g. Portacath TM) ............ 9
Implementation.......................................................................................................................... 9
Related Policies, Procedures, Guidelines and Legislation .......................................................... 9
References ................................................................................................................................ 10
Definition of Terms................................................................................................................... 10
Search Terms ............................................................................................................................ 12
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
1 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
Purpose
To provide information to Canberra Hospital and Health Services staff on the correct, safe
and appropriate collection of blood cultures to confirm and identify the possible causative
micro-organism in bacteraemia / septicaemia.
Alerts
Scope
1.
2.
Blood culture collection is an standard Aseptic Non Touch Technique.
For Adult patients –One set of adult blood culture bottles consists of two separate
bottles (one aerobic and one anaerobic – (see Figure 1).
3.
For each septic episode two complete separately drawn sets of blood cultures are
required (four bottles in total)
For paediatric patients - A single bottle system is used (see Figure 2). One set (one
bottle) is required unless the patient is considered to be immunocompromised in which
case two sets (two bottles) are required to be collected.
4.
5.
6.
7.
8.
9.
Paediatric bottles can be used for adult patients when only a small blood volume is able
to be collected.
Skin preparation is with 2% Chlorhexidine in 70% alcohol skin preparations and must be
allowed to dry for skin disinfection to be effective.
A vacuum container set with a safety lock scalp vein needle is to be utilised to minimise
the risk of occupational exposure and to facilitate appropriate specimen collection.
Patients with a centrally inserted vascular access device (CVAD) require one set of blood
cultures collected via venepuncture from a peripheral site and two sets from two
separate lumens of the CVAD to assist in diagnosis of systemic infection versus CVAD
related infection.
When a micro-organism is identified and the patient is receiving the appropriate
antibiotics it is unnecessary to collect any further blood cultures unless:
a. The patient is still febrile after 48 hours of treatment of the appropriate agent; or
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
2 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
b. The patient becomes febrile after a period of being afebrile
c. The patient has Staphylococcus aureus bacteraemia, Candidaemia or infective
endocarditis. These cases require serial collection until they are culture negative.
Paediatrics
If you are inserting an IV cannula in a neonate, infant or child and blood cultures collection is
indicated, the blood cultures can be taken from the cannula at the time of insertion,
providing the cannula has been inserted under strict standard Aseptic Non-Touch Technique
(ANTT). Specimens collected in this manner have a high risk of contamination.
Back to Table of Contents
Scope
This document pertains to all patients that require collection of blood cultures at the
Canberra Hospital and Health Services.
This document applies to:
 Medical Officers
 Nurses and Midwives who are working within their scope of practice
 Phlebotomists (venepuncture only, not intravascular device access)
Indications for collection of blood culture:
 Clinical conditions suggestive of microorganisms in blood e.g. fever, chills, rigors,
tachycardia, tachypnoea.
 Fever and hypotension not explained by non-infectious causes.
 Fever in the presence of neutropenia and when IV devices are present.
 In the absence of fever:
o Patient with focal infection e.g. pneumonia, meningitis, osteomyelitis.
o Neonates and infants with unexplained hypothermia.
o Children or elderly with sudden failure to thrive.
o Elderly with deterioration from baseline status that is confusion, sudden altered
mentation or frequent falls.
o Patient with renal insufficiency and unexplained leucocytosis altered mentation or
unexplained tachycardia.
o Unexplained pulmonary, renal, or hepatic dysfunction in an immuno-compromised
or critically ill patient.
o Unexplained haemodynamic instability.
 Ongoing monitoring of Staphylococcus aureus bacteraemia, Candidaemia, and infective
endocarditis.
Back to Table of Contents
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
3 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
Section 1 – Peripheral blood culture collection
Equipment
 Patient pathology identification labels x 4 (if available) (or new pathology system)
 Pathology collection form
 Dressing trolley
 Alcohol based hand rub (ABHR)
 Alcohol swabs or alcohol 70% with chlorhexidine swabs x 4
 3ml Chlorprep (2% chlorhexidine and 70% alcohol) x 1
 Detergent impregnated wipes
 Basic dressing pack x 2
 Vacuum container system – 2 sets
o Blood transfer device
o Safety lock - scalp vein set 21G
 Pressure dot / bandaid
 Sterile gloves
 Bactec Blood culture bottles
 Adults Aerobic blood culture bottles x 2
 Adults Anaerobic blood culture bottles x2
o Paediatric
 Paeds Plus/F Plus bottle
 Protective glasses or goggles
 Biohazard sharps container
 Clinical waste receptacle
 Tourniquet
 Under pad
Procedure
1. Perform hand hygiene.
2. Collect all equipment required (including personal protection equipment) and place on
bottom of trolley that has been cleaned from top to bottom with detergent‐based
wipes. Bring trolley to the patient zone.
3. Ensure that the relevant history and tests are stated on the pathology blood culture
request form as this may affect incubation requirements.
4. Check expiry date for each bottle and mark 10 mL above the broth for fill level for adult
patients (8 to 10 mLs) and 3 mL above for paediatric patients.
5. Check patient identification and where possible, inform patient of the procedure and
gain verbal consent.
6. Open dressing pack and place blood collection equipment and skin preparation onto
sterile field. Bloods culture bottles should be placed to edge or on back corner of sterile
field.
7. Prepare winged infusion set, blood transfer device, and other equipment.
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
4 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
Remove the cap of each blood culture bottle and scrub the vial stoppers and neck of
bottle well using alcohol 70%, or alcohol 70% with chlorhexidine, and allow to dry
completely. Place to side or on back corner of sterile field.
Perform hand hygiene (Moment 1).
Position patient appropriately, apply tourniquet to palpate and identify appropriate
vein.
Perform hand hygiene (Moment 2) before the procedure.
Put on sterile gloves.
Using Chloraprep (2% chlorhexidine and 70% alcohol) disinfect venepuncture site with
gentle back-and-forth strokes, progressing from the insertion site to the periphery for
30 seconds, allow to dry for a further 30 seconds.
It is preferable that the venepuncture site is not re-palpated after skin preparation
however if necessary, re-palpate the venepuncture site to check vein position with
sterile gloves that have not been contaminated.
Perform venepuncture using winged infusion set and syringe or vacuum container.
Release tourniquet during procedure where appropriate, this will contaminate the
gloved hand and having an assistant to do this or by using a sterile towel or the
non‐dominant hand is advised.
Inoculate the blood culture bottles aerobic bottle (blue) first followed by anaerobic
(purple) bottle.
Place 10 mL blood per bottle (20 mL/set), keeping blood culture bottle upright and
at/below the level of the venepuncture.
If blood collected via a syringe use a blood transfer device to transfer 8 to 10mls of
blood into each bottle – aerobic followed by anaerobic
If collecting additional blood pathology tubes always collect/inoculate the blood culture
bottles FIRST.
Remove winged infusion set and apply cotton ball and pressure to site (where possible
obtain patient assistance to hold and apply pressure).
Invert bottles gently several times to prevent clotting.
Discard sharps, collect all rubbish/dirty items and dispose of appropriately.
Immediately after specimen collection at the patient bedside collector must personally
label each bottle with patient name, medical record number (MRN), date/time for
collection of blood and location of site used. If using pre printed patient pathology
identification labels do not cover any bar codes or the bottom of the bottle.
Place bottles into biohazard bag and arrange to send to the lab with request form.
Transport bottles at room temperature. Do not send more than two blood culture
bottles at a time in the pathology pneumatic rapid transport tube system, due to the
potential for breakage.
Remove gloves and perform hand hygiene (Moment 3) after the procedure.
Repeat entire procedure for second set of blood cultures collected from a different
peripheral site, using new dressing pack and general aseptic field and maintaining
standard aseptic technique. Ideally if time permits collect second set 30 minutes later. If
patient is acutely unwell and requires empirical antibiotics collect second set as soon as
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
5 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
possible. If original specimen has been collected by phlebotomist, second set should
ideally be collected by a different collector if available.
Blood is normally sterile; the purpose of collecting blood cultures is to identify any bacteria
or fungi growing in the blood that may cause sepsis. If there are pathogens growing in the
blood they take time (usually up to 48 hours) to grow and to be identified in the laboratory.
Explain to patient that results may not be available for 48 hours or more. Once cultures have
been collected do not delay administering antibiotics if they have been ordered, do not wait
for culture results as any delay in antibiotic administration may be detrimental to the
patient.
Documentation
Place small peel off barcode label from blood culture bottles in clinical notes and document:
(a) number of sets of blood cultures taken,
(b) site/s of collection and
(c) reason for site choice if this differs from a peripheral site.
Additional Samples
1. Additional sample may be requested by the Medical Officer.
2. A peripheral vein (upper limb) access is preferable, however if the patients venous
access is poor, one set may be taken from a central line, and clearly marked as such on
bottles.
3. At least one set of blood cultures should be obtained peripherally.
4. There is usually no need for more than three sets of blood cultures to be collected
during any single febrile event.
5. Repeat blood cultures should also be collected routinely in Staphylococcus aureus
bacteraemia and candidaemia, to determine when cultures become negative on
appropriate antibiotics.
6. Patients with suspected or confirmed infective endocarditis should have further regular
cultures to determine when cultures become negative.
7. Specific blood culture bottles are required for Mycobacterial species. These also are
enhanced to support the growth of fungi in patients at high risk for fungaemia. Contact
the ACT Pathology to organise Mycobacterial bottles.
8. In an extremely ill patient, additional blood cultures may be taken sequentially from two
separate sites prior to commencing antibiotic treatment.
Back to Table of Contents
Section 2 – Collection from a Central Venous Catheter (CVC) or Peripherally
Inserted Central Catheter (PICC)
Patients with a Central Venous access device (CVAD) also need to have one set of blood
cultures, collected peripherally by venepuncture, to assist in determining diagnosis of
systemic infection or CVAD related infection.
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
6 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
Ongoing blood culture specimens can be collected from CVAD only.
Equipment
 Patient pathology identification labels x 4 (if available)
 Dressing trolley
 Alcohol based hand rub (ABHR)
 Alcohol swabs or alcohol 70% with chlorhexidine swabs x 4
 2% chlorhexidine in 70% alcohol skin preparation swabs x 4
 Detergent impregnated wipes
 Basic dressing pack x 2
 Syringe 20 ml x 2 for blood collection

Syringe 10 ml x 2 for Saline flush solution or premix Saline flush solution x 2
 Saline flush solution or premix Saline flush solution x 2
 Vacuette blood transfer device x 2 sets
 Replacement Needleless injection caps or positive pressure valves
 Sterile gloves
 Bactec Blood culture bottles
o Adults
 Aerobic blood culture bottles x 2
 Anaerobic blood culture bottle

o Paediatric
 Paeds Plus/F Plus bottle
 Protective glasses or goggles
 Biohazard sharps container
 Clinical waste receptacle
 Tourniquet
 Under pad
Procedure
1. Perform hand hygiene.
2. Collect all equipment required (including personal protection equipment) and place on a
trolley cleaned with detergent‐based wipes and bring to the patient zone.
3. Request assistance from another staff member.
4. Ensure that the relevant history and tests are stated on the blood culture request form
as this may affect incubation requirements.
5. Check expiry date for each bottle and mark 10 mL above the broth for fill level for adult
patients and 3 mL above for paediatric patients.
6. Check patient identification and inform patient of the procedure and its purpose.
7. Open dressing pack and place blood collection equipment and skin preparation onto
sterile field.
8. Remove the cap of each blood culture bottle and scrub the vial stoppers and neck of
bottle well using alcohol 70%, or alcohol 70% with chlorhexidine, and allow to dry
completely. Place to side or on back corner of sterile field.
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
7 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
9. Perform hand hygiene (Moment 1).
10. Position patient appropriately for ease of access to CVAD.
11. Perform hand hygiene (Moment 2) before the procedure.
12. Don sterile gloves.
13. Prepare syringe, blood transfer device, and other equipment.
14. Ask assistant to attend hand hygiene, don non sterile gloves and lift CVAD lumens.
15. Place sterile drape under CVAD lumens.
16. Check needleless injection cap for visible contamination and if present change
needleless injection cap prior to collecting blood cultures.
17. Swab the needleless injection cap thoroughly with 2% chlorhexidine in 70% alcohol
swab for 10 seconds. Allow to dry for 30 seconds.
Note
When collecting blood cultures from a CVAD Do Not discard blood or fluid as per
routinue practice for CVAD blood draws. Pathogens may be residing within the lumen
and collecting the initial blood draw will lead to greater accuracy.
Syringe draw
18. Attach syringe to injection site and withdraw 20 mls of blood into syringe and
disconnect syringe from CVAD.
19. Attach blood transfer device to syringe and inoculate the blood culture bottles aerobic
bottle (blue) first followed by anaerobic bottle.
20. Invert bottles gently several times to prevent clotting.
Blood transfer device collection
21. Attach luer lock access device (blue top) device to CVAD injection site.
22. Connect the blood cultures bottles to the transfer device and inoculate the blood
culture bottles aerobic bottle (blue) first followed by anaerobic bottle.
23. Place 10 mL blood per bottle (20 mL/set), keeping blood culture bottle upright and
at/below the level of the CVAD.
24. Invert bottles gently several times to prevent.
25. Procedure Continuation for both methods
26. If collecting additional blood pathology tubes always collect/inoculate the blood culture
bottles FIRST.
27. Flush CVAD lumen with 20 mLs 0.9% sodium chloride for injection using a pulsatile
action.
28. Replace needleless injection cap as per Central Venous Access Device (CVAD)
Management – Children, Adolescents and Adults (NOT Neonates) procedure if there is
residual blood visible within device.
29. Discard sharps, collect all rubbish/dirty items and dispose of appropriately.
30. At the patient bedside label each bottle with patient name, MRN, date/time for
collection of blood and location of site used for each set (eg. Blue lumen PICC). Do not
cover any bar codes or the bottom of the bottle.
31. Place bottles into biohazard bag and arrange to send to the lab with request form.
Transport bottles at room temperature.
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
8 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
32. Remove gloves and perform hand hygiene (Moment 3) after the procedure.
Repeat entire procedure for 2nd set of blood cultures from a different lumen, using new
dressing pack / sterile field and maintaining standard aseptic technique. If original
specimen has been collected by phlebotomist, second set should ideally be collected by a
different collector if available.
Back to Table of Contents
Section 3 – Collection from an Implanted Venous Access Devices (e.g.
Portacath )
TM
Patients with an Implanted Venous access device also need to have one set of blood cultures
collected by venepuncture to assist in determining diagnosis of systemic infection or device
related infection.
For ongoing testing only Implanted Venous Port specimens are required.
Procedure
1. The implanted venous device should be accessed as per the Central Venous Access
Device (CVAD) Management – Children, Adolescents and Adults (NOT Neonates)
procedure
2. For blood culture collection from implanted device follow steps in section 2 above
Back to Table of Contents
Implementation
This procedure will be communicated to relevant staff via:
 organisation wide notification of updated procedure
 discussion at clinical / ward based meetings
 incorporation into existing education and training programs
 blood culture collection posters available in clinical areas.
Back to Table of Contents
Related Policies, Procedures, Guidelines and Legislation
Policies
CHHS Clinical Policy Patient Identification and Procedure Matching
Procedures
CHHS Standard Operating Procedure Central Venous Access Device (CVAD) Management –
Children, Adolescents and Adults (NOT Neonates)
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
9 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
CHHS Standard Operating Procedure Venepuncture Blood Specimen Collection
CHHS Standard Operating Procedure – Aseptic Non Touch Technique
CHHS Standard Operating Procedure – Patient Identification: Pathology Specimen Labelling
Guidelines
NSW Health Clinical Excellence Commission (CEC), Sepsis Kills – Blood Culture collection
Guideline http://www.cec.health.nsw.gov.au/programs/sepsis
Back to Table of Contents
References
Burton, G & Engelkirk P. G., (2000) Microbiology for the Health Sciences 6th
Edition, Lippincott Williams & Wilkins, p285- 286.
BACTEC Blood Collection Instructions, Becton Dickinson Pty Ltd, NSW Australia.
https://www.bd.com/ds/technicalCenter/charts/ch_3_2340.pdf
NSW Health Clinical Excellence Commission (CEC) 2013, Sepsis Kills Program: Adult blood
culture sampling guideline accessed 01/07/2015
http://www.cec.health.nsw.gov.au/programs/sepsis
Mandell, Douglas and Bennett’s principles and practices of infectious diseases – 7th
edition (2010). Edited by Gerald Mandell, John E Bennett & Raphael Dolin, Churchill
Livingstone Inc. pp235 -236
Mylotte JM., Tayara A., (2000) Blood Cultures: Clinical Aspects and Controversies.
European Journal of Clinical Microbiology and Infectious Diseases 19: p157 – 163.
Wenzel, R., (1997) Prevention and Control of Nosocomial Infections 3rd edition.
Williams and Wilkins A, Waverly Company.
World Health Organization (2010), WHO guidelines on drawing blood: best practices in
phlebotomy, accessed 01/07/2015
http://whqlibdoc.who.int/publications/2010/9789241599221_eng.pdf
Back to Table of Contents
Definition of Terms
Aerobic: Occurring in the presence of oxygen or requiring oxygen to live.
Alcohol Based Hand Rub (ABHR): Alcohol containing preparation designed for application to
the hands in order to reduce the number of viable micro organisms with maximum efficacy
and speed.
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
10 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
Anaerobic: Occurring in the absence of oxygen or not requiring oxygen to
live. Anaerobic bacteria grow without the presence of oxygen.
Aseptic Non Touch Technique (ANTT): - ANTT is a technique used to prevent contamination
of key parts and key sites by microorganisms that could cause infection. In ANTT, asepsis is
ensured by identifying and then protecting key parts and key sites by hand hygiene, nontouch technique, using new sterilised equipment and/or cleaning existing key parts to a
standard that renders them aseptic prior to use
Candidaemia - is a bloodstream infection caused by yeast (a type of fungus) called Candida.
These infections often result in long hospital stays, high medical costs, and poor outcomes
Central Venous Catheter (CVC) - Central Venous Catheter is an intravascular device placed
into a large vein in the neck (internal jugular vein), chest (subclavian vein) or groin (femoral
vein) with the tip terminating in the superior vena cava or right atrium. CVC’s can be non
tunnelled (percutaneous) or tunnelled; inserted with a tunnel under the skin creating
distance between skin insertion site and vascular insertion site (Hickman TM, VascathTM).
Contamination / False positive - A false positive is defined as growth of bacteria in the blood
culture bottle that were not present in the patient’s bloodstream and were introduced
during sample collection. Contamination can come from a number of sources: the patient’s
skin, the equipment used to take the sample and transfer it to the culture bottle, the hands
of the person taking the blood sample, or the general environment.
Fungaemia - a bloodstream infection caused by fungi.
Hand Hygiene (HH): A process that reduces the number of micro-organisms on hands.
Hand hygiene is a general term applying to the use of soap solution (non-anti-microbial or
anti- microbial) and water or water-less antimicrobial agent to the surface of the hands
(e.g. alcohol based hand rub).
Implanted Venous Access Port (Port, PortacathTM, PasportTM) - an implantable medical device
consisting of a fluid reservoir connected to a central venous catheter. The device is
completely underneath the skin and the catheter tip generally terminates in the superior
vena cava. Access is made through the skin into the reservoir using a non-coring needle
device (Gripper™)
Peripherally inserted Central Catheter (PICC) – A PICC is an intravascular catheter inserted
in a peripheral vein in the arm, such as the cephalic vein, basilic vein or brachial vein, and
then advanced proximally toward the heart until the tip rests in the distal superior vena cava
or cavoatrial junction.
Back to Table of Contents
Doc Number
CHHS16/004
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Area Responsible
Page
Infection Prevention
11 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register
CHHS16/004
Search Terms
Blood culture, Bacteraemia, Bloodstream Infection, Central Venous Catheter, CVC,
Fungaemia, Infection, Peripherally inserted Central Catheter, PICC, Sepsis, venepuncture
Back to Table of Contents
Disclaimer: This document has been developed by ACT Health, <Name of Division/ Branch/Unit> specifically for
its own use. Use of this document and any reliance on the information contained therein by any third party is at
his or her own risk and Health Directorate assumes no responsibility whatsoever.
Date Amended
6 June 2017
Doc Number
CHHS16/004
Section Amended
Minor Amendment to skin
prep used in section 1
Version
1
Issued
01/02/2016
Review Date
01/02/2019
Approved By
CHHSPC
Area Responsible
Page
Infection Prevention
12 of 12
and Control
Do not refer to a paper based copy of this policy document. The most current version can be found on the ACT Health Policy Register