Biosafety in the Mycobacteriology

Laboratory
June 2011

Natalia Shubladze, TB lab Consultant

Lynee Galley, Senior Program Specialist, International
Affairs, LabCap
 Part IY
Waste
management
 Waste management

All materials used for TB diagnostics should

be treated as infectious waste.
Disposable materials should be autoclaved
or incinerated
Reusable materials (e.g. forceps, transport
trays, glassware) should be disinfected using
appropriate disinfectants (e.g. 5% phenol)
before autoclaving and washing.
 Decontamination
Decontaminate specimens and cultures by one
of the following methods:

Liquid decontamination using disinfectants

Autoclaving
Incineration to ash in oven

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 Autoclave
 Moist heat, in the form of saturated
steam under pressure, is the most
effective and reliable means of
destroying all forms of microbial life
(CDC,OHS)
 Steam must circulate, penetrate,
and contact all surfaces for efficient
decontamination
 Tubercle bacilli are killed by steam
heat
 If containers are closed, water
should be added to generate steam
within the container
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Proper use of the Autoclave
Review and understand the owner’s manual
before using any autoclave
Contact the manufacturer to request on-site
training as needed
Do not autoclave items containing corrosives
(e.g. acids, bases, phenol), solvents or volatiles
(e.g. ethanol, methanol, chloroform) or radioactive
materials
Do not use bleach as a disinfectant for waste that
is autoclaved- chlorine gas is generated and
autoclave will be corroded.
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 Autoclave Run Settings
 Materials for decontamination:
-132°C (115 kPa), for 30 minutes

 To sterilize media and materials:

– 121°C (115 kPa), 20 minutes
Notes:
Disposable and non-disposable materials
should not be autoclaved in the same
container
Clean and dirty material should NOT be
autoclaved together 7
 QC for Autoclave
 Add indicator tape each time
autoclave is run
-Tape changes colour when correct
temperature and time are achieved

 Weekly check autoclave function

– Spores of Bacillus stearothermophilus
ATCC 1953 on a strip or in an ampule
– Spores should not germinate and grow
when placed in broth or incubated after
autoclaving
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Disposal of decontaminated waste

 Burying according to country or local regulations

 Commercial waste and chemical management
companies may pick up decontaminated waste
 Other means according to country or local
regulations

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Remember SAFETY FIRST!
• Skills
• Accountability
• Facility
• Equipment
• Training
• YOU
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 Literature
• CDC/NIH Biosafety in Microbiological and Biomedical Laboratories, 5th
Edition. (2007) http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm
• Laboratory biosafety manual. 3rd edition, 2004. WHO.
http://www.who.int/csr/resources/publications/biosafety/en/
• Biosafety7.pdf Biosafety recommendations for the contained use of M.
tuberculosis complex isolates in industrialised countries. P Herman et al, 2006.
Scientific Institute of Public Health, P Herman et al, 2006. Scientific Institute of
Public Health, Brussels, Belgium.
http://www.biosecurite.be/CU/PDF/Mtub_Final_DL.pdf
• Interim Laboratory Biosafety Guidance for Extensively Drug-Resistant (XDR)
M. tuberculosis strains.
http://www.cdc.gov/tb/XDRTB/BiosafetyGuidance_xdrtb.htm
• Laboratory Biosafety Levels for TB Diagnostics (African Center for Integrated
Laboratory Training)
• Thomas M. Shinnick, Ph.D. Mycobacteriology Laboratory Branch Division of
Tuberculosis Elimination Biosafety in the Mycobacteriology Laboratory