University of Cambridge
NIHR BRC Phenotyping Hub, Department of Medicine
Risk Assessment Form
This form is to be filled in if hazard ratings/quantities/dilutions make University procedures and/or
“Good Laboratory Practice” insufficient control.
Group leader: Anna Petrunkina Harrison
Personnel involved: Simon McCallum, Chris Bowman, Esther Perez, Renato Daniel dos Reis
Colaco, Natalia Savinykh and all users of flow cytometry equipment in the analyser room
Activity being assessed:
HANDLING AND ANALYSING SAMPLES from
UNSCREENED HUMAN tissue and blood
Hazards identified:
Unscreened human blood and other unscreened tissues may contain or be contaminated with
Blood Borne Viruses including HIV, Hep BV, Hep CV amongst others or harbour other
agents associated with them and should be handled as such.
Hazards from contamination include:
1. Percutaneous inoculation from sharps, including needles, scalpels, broken glass etc.
including contamination of exposed skin or pre-existing cuts and abrasions etc.
2. Inhalation of aerosols from shaking, agitation, mixing and failure of containment etc.
3. Ingestion from inadvertent sample to mouth transmission.
4. Contamination of Conjunctivae by inadvertent contact with eyes.
resulting in infection.
Therefore, unscreened human samples should be handled at Containment Level 2 in accordance with
the revised ACDP guidance from the HSE.
If there is a significant risk that, or it is actually suspected that, the blood/tissue is infected
with any Blood Borne Viruses (BBVs), including the above, or any other pathogenic agent,
then a separate Risk Assessment must be carried out in order to reduce the risk (e.g. fixation
with PFA) and an appropriate Containment Level used. Such samples can be processed in the
analyser room only after enquiries with Biological Safety Officer and after approval by
Departmental Biosafety Committee.
Guidance Documents: See
University booklet “Safe Biological Practice (SBP) – for Prevention and Control of Exposure to
Biological Agents in the Laboratory”
University Clinical School Guidance on Blood Taking, Blood Handling and Work at Containment
Level 2.
Advisory Committee on Dangerous Pathogens (ACDP) Guidance “Protection against blood-borne
infections in the workplace- HIV and Hepatitis (HSE/HMSO : ISBN 0 11 321953 9)
Revised Advisory Committee on Dangerous Pathogens Guidance (www.doh.gov.uk/ACDP):-
Control measures to reduce the level of risk:
Unscreened human samples processed for flow cytometry analysis should be handled at
Containment Level 2. In addition to the measures and facilities normally specified for work at
Containment Level 2 (see relevant guidance), the additional measures listed below should also
apply when, as here, Containment Level 2 is specified.
These extra precautions should be in constant use to guard against percutaneous inoculation, contamination
of the skin, conjunctivae, mucous membranes and the working surfaces.
• All work must be conducted in a designated containment level 2 work area (Phenotyping Hub analyser lab
space) with sufficient space to work safely. The right hand area in the analyser room where flow cytometers
are located is designated as such an area. The left hand area is office and storage area operated at
Containment Level 1 – no samples can be placed on these benches.
• Access to CL2 areas is restricted to authorised staff ONLY, in order to ensure that the person(s)
conducting the work is (are) free from the risk of disturbance or accidental contact with others. Access is
operated via personal ID cards, and transfer/borrowing these cards is not permitted. All users will be advised
on the conversion of the analyser area into a CL2 workspace, and on the necessity to adhere to procedures
relevant for handling their own samples and to perform risk assessment for their own experiments. This
advice is now incorporated in the training for flow cytometry area.
• The use of sharps and glassware must be avoided in the Analy room. Where such use is essential, particular
care must be taken in their handling and disposal after contact with unfixed materials, and any exception can
be only subsequent to the application and review of the RA for the protocols.
• Lesions on exposed skin should be covered with waterproof dressings.
• Protective clothing must include an appropriate dark blue CL2 dedicated laboratory gown, disposable
gloves and other personal protective equipment appropriate to the task, including eye protection when
appropriate. It must be stated in the researchers project RA’s which PPE and when has to be worn.
• The workstations must be kept clear of unnecessary equipment and not used for storage.
• When operating analysers, samples must be handled with care, especially when taking them off the
injection port. As all flow cytometers operate at a higher than ambient pressure, aerosols are generated when
sample tubes are exchanged. These aerosols can neither be contained nor prevented.
THEREFORE, NO UNSCREENED SAMPLES WHICH ARE NOT PFA-FIXED CAN BE
PROCESSED IN FLOW CYTOMETRY LABORATORY WITHOUT PRIOR REVIEW OF
PROJECT RISK ASSESSMENTS WITH RESPECT TO EVALUATING RISKS RELATED TO
PROPAGATION OF INFECTION BY AEROSOLS ACCORDING TO THE PROCEDURE
DESCRIBED BELOW:
Investigators with unscreened samples which are not PFA-treated must seek advice (prior starting their
experiments and using flow cytometry equipment) from Biological Safety Officer. Dr. Mark Wills will
assess whether the risk assessment for experiments is adequate, in particular, whether the risk has been
sufficiently reduced by additional procedures and factors (e.g. patient risk group, cell isolation, depletion,
fixation, viral inactivation, screening for BBV) and refer to the Safety Committee in cases of enhanced risk.
In situation where unfixed samples are handled in flow cytometry room and the residual risk is considered to
be above low levels, and an approval of Safety Committee cannot be obtained, samples cannot be processed
on analysers but the users are advised to contact Hub staff for advice on using operator service for analysing
the samples on sorter placed in MSC.
• The bench surface and any equipment must be disinfected immediately after completion of work
Following disinfection policy is in operation:
1) After completion of flow cytometry work, flow cytometric equipment must be cleaned as described
in the guidance notes (detergent, followed by water). The waste tank needs to be supplemented with
Virkon tablets in the quantity prescribed for each particularinstruments (final concentration of ~1%).
NOTE: Virkon must be never used for cleaning equipment!
2) In case of spillages, surface area has to be sprayed with Trigene and wiped.
• For all staff entering the flow cytometry area for conducting work, Hep BV vaccine, together with followup procedures, should be considered.
• Contaminated waste will be disposed of in accordance with local guidance and rules for the safe
disposal of Containment Level 2 waste; including chemical sterilisation of liquids with autoclaving
and incinerating for solids as appropriate. If it can be carried back in containment,users should feed
it in to their waste streams.
Level of risk remaining:
Residual risk of percutaneous inoculation; which must be minimised by the avoidance of sharps.
Residual risk of splashes in eyes; which must be minimised by use of suitable eye protection.
Emergency procedures:
NB: See Guidance Documents as above.
First Aid: Wash any contaminated skin, conjunctivae or mucous membrane immediately.
In the event of a wound, it should be allowed to bleed by irrigation under running water.
Seek medical Advice and contact University Occupational Health Service immediately.
Spillage: All spillages and surface contamination must be immediately cleaned up and removed
including decontamination with a suitable validated disinfectant (10% solution of Trigene).
Blockages of the equipment: If an analyser, Hub staff MUST be advised immediately on the nature
of the sample, and deblocking procedure must be performed under standard GLP (with syringe and
tubing to inject decon/trigene or ultrasonic bath; for non-fixed samples after shutting down the
fluidics/depressurizing the system and in suitable working area covered by separate RA and SOP for
deblocking procedure).
Name and status of assessor:
Anna Petrunkina, Director of the Cell Phenotyping Hub
Date of assessment:
01/09/2014
Signature of assessor:
Revision due date:
01/09/2015