STANDARD OPERATING PROCEDURE
Procedure:
School/Department:
SOP prepared by:
Working with risk group 2 microorganisms
School of Biological Sciences
Nick Coleman (SMB) edit M Joseph, N Firth
Section 1 - Personal Protective Equipment
1. Lab coat or gown. (Gown is preferable for PC2 work)
2. Long hair tied back.
3. Proper enclosed shoes.
4. Safety glasses / goggles if splash/squirt/spray risk present.
5. Nitrile or latex Gloves
Section 2 – Potential Hazards + Safety precautions
1. See corresponding risk assessment. Briefly, hazards are infection of lab worker,
infection of other people, infection of plants or animals, and escape of the
microorganism into the environment. These hazards are significant for risk group 2
microbes. The degree of risk will depend upon the individual organism, the route
of exposure (eg. inhaled, ingested, in a wound, in the eye), and the number of
organisms involved.
Section 3 – Procedure
1. Work in a PC2-approved laboratory facility (see AS 2243.3 for details)
2. Know the locations of emergency equipment, especially eyewash, the locations
of emergency exits, and the emergency evacuation procedure.
3. Know the risks of the organisms you are working with, eg. check MSDS if
available, or ask your supervisor
4. Worker should be previously trained in microbiological methods, especially
aseptic technique. If untrained (eg. undergraduate students), the worker should
be closely & constantly supervised by someone with microbiological training.
5. Worker should be familiar with the Australian Standard document AS 2243.3,
which describes appropriate work practices in a microbiology lab. This
document must be readily available in all microbiology labs.
6. Worker must be aware of any pre-existing relevant health conditions, and inform
their supervisor of these, if these would impact on microbiological work. eg.
Immunocompromised status due to HIV infection or medication would make the
worker much more susceptible to infection.
7. Never eat, drink, smoke or apply cosmetics in the microbiology lab. Never put
anything in your mouth (eg. don’t chew the end of pens).
8. Do not use your mobile phone while doing microbiological lab work- this can act
as a vector for carrying microbes outside the lab.
9. Work bench should be clear of clutter and personal possessions (eg. no bags,
clothing etc). Only the equipment necessary for the tasks to be performed
should be present on the bench. Do not sit on the lab bench.
10. Bench surface should be disinfected before beginning work, and after
completion, with 80% (v/v) ethanol solution. It is good practice to also similarly
sterilise any tools required (eg. the surface of Pipetman-type pipettes).
Microbiological loop is sterilised immediately before and after each procedure
by flaming in bunsen burner. Glass spreader is sterilised immediately before each
procedure, and after, by dipping in 100% ethanol and briefly flaming (see risk
assessment for Bunsens and Spread-plating). All other equipment + media must
Issue date: 20/11/12
Review date: 20/11/13
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be sterilised by autoclaving or filtration.
11. Turn Bunsen burners off when not actively doing lab work requiring these. This
helps keep the lab cool, and minimises the fire hazard. Do not leave a Bunsen
flame unattended, even for a few minutes.
12. When making transfers of cultures, streaking plates etc, work close to the Bunsen
flame – this minimises deposition of airborne microbes onto/into media.
13. Work in a biosafety cabinet for any manipulations of risk group 2 organisms that
are likely to generate aerosols – this includes vortexing, sonicating, pipetting,
centrifugation, and opening tubes. See SOP for biosafety cabinet. Note that a
laboratory gown (not coat) must be worn when using a biosafety cabinet.
14. All cultures must be labelled with the name of the organism (species and strain),
the name or initials of the worker, and the date. Cultures must be incubated in
the appropriate location, and must not be taken out of the lab.
15. Risk group 2 cultures must be transported inside a secondary unbreakable
container if they need to be transported outside the PC2 lab. A strong plastic
box with a lockable lid is ideal. See also SOP for Transport of GMOs.
16. Risk group 2 cultures must be stored in a secondary unbreakable container, and
be properly labelled (as above), and records must be kept of the cultures. Also
see SOP: storage of GMOs
17. Upon finishing work, sterilise the bench again, and discard all unwanted cultures
into the appropriate location. All microbial cultures must be destroyed by
autoclaving, they cannot be put down the sink, or in the regular garbage. See
also the SOPs for Disposal of GMO contaminated waste and Disposal of
Biological waste.
18. Wash your hands with antiseptic soap before leaving the laboratory, eg. with
soap containing chlorhexidine.
19. Remove your lab coat or gown before leaving the laboratory. Lab coats or
gowns used in PC2 labs must not leave the facility, unless they are autoclaved
first. Autoclaving must be done before laundering the coat/gown.
Section 4 – Spills / Incidents
1. Report all spills and other incidents to your supervisor, and if a serious incident
occurs, this needs to be formally reported via the online reporting system.
2. In the case of a large spill of a risk group 2 organism (eg. >100 ml), allow 20 min
for aerosols to settle before attempting to clean up the spill. Alternatively, wear a
full-face respirator (found in spill kit) if you need to clean it immediately.
3. Contain the spill by surrounding with paper towel or an absorbent ‘boom’ from
the spill kit, or other absorbent material
4. Disinfect the spill by adding freshly prepared bleach solution containing 1%
hypochlorite. Allow at least 10 min for the bleach to kill the microbes.
5. Soak up all the liquids with paper towels, and dispose of all the towels and any
other spill containment items used into a strong plastic bag (eg. a ‘wet garbag’
or a biohazard bag).
6. This bag containing the spill materials should NOT be autoclaved, as bleach will
corrode the autoclave. Instead, this should be disposed of as medical waste, or
as hazardous chemical waste.
Section 7 - References
1. Australian Standard document AS 2243.3
2. Risk assessment and SOP: Disposal of biological waste
3. Risk assessment and SOP: Biosafety cabinet
Issue date: 20/11/12
Review date: 20/11/13
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4. Risk assessment and SOP: Biohazard spills
SOP Training Confirmation
By signing below, these individuals confirm that they have read and
understood the SOP, and agree to always follow the instructions in this SOP
when performing this procedure.
Position
Name
Signature
Date
Supervisor
employee / student
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WHS Committee Approval
Representative: A. Prof Frank Seebacher Chair Safety Committee
Signature: ........................................................ Date: .....................................
Issue date: 20/11/12
Review date: 20/11/13
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