LABORATORY FACILITIES AND SAFETY
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Problem Scenario
Your laboratory is specialized in virology. There has been an important epidemic of H5N1 in your country and the Ministry of Health has asked you to be the referent laboratory for this pathology. What must you do to ensure biosafety in your laboratory?
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The Quality System
Process Control
(Quality Control & Specimen
Management)
Purchasing & Inventory
AssessmentOccurrence
Management
Information Management
Process Improvement
Customer Service
Facilities & Safety
Organization Personnel Equipment
Documents & Records
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Learning Objectives
At the end of this activity, you will be able to: Discuss the importance of laboratory design Know what, where and when the risks are Discuss the importance of using appropriate
biosafety equipment Describe the four biosafety levels Outline factors that need to be considered
when assessing risk Keep appropriate biosafety documentation
1. Importance of laboratory design
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Laboratory design
1-Path followed by the sample: Reception and registration of patients Sampling rooms Dispatch between different laboratories Analysis of samples
Results Delivery and filing of results Related services: Secretariat, Offices,
Washery, Preparation and sterilization, Storage, Cold room.
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Premises
Reception : > reception and registration of patients > reception and registration of samples from
other departments
Sampling rooms > samples and sorting of samples
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Blood clotting
Haematology
Biochemistry
Washroo
m
Reception and
administration
Bacteriology
Gynaecological specimens
Blood specimens
Common room, stairs
to offices
TOTAL AREA 104 M2
ENTRANCE
Disinfection
Plan
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Washery with autoclaves for sterilization, wash and dry of glassware
Preparation, sterilization and distribution of culture media and reagents
Stock room Cold room Culture room others
Service rooms
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Premises
High ceiling Walls and ceiling with glossy paint :
easy to clean and disinfect. Floor easy to clean and disinfect
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Access to premises
Only authorized persons: technicians, biologists, and maintenance staff ( badges)
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Safety during serviceNo unauthorized persons
No friendsNo childrenNo animals
PleaseOPENAND
CLOSE
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Premises cleaning
Maintenance on a daily basis: Benches Floor
Maintenance on a weekly basis: Ceiling and walls
Maintenance on a weekly basis: Closets Fridges
Date and name of person
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Benches
Non porous covering, easy to clean and resistant to chemicals and disinfectants (Chlorine, etc.)
No wood, no steelGlass, ceramic tiles, etc.
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2. Know the risks
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The risks Most Frequently Reported Infections in US, 1979-1999
Disease or Agent No. of Cases
M. tuberculosis 223
Q fever 176
Hantavirus 169
Hepatitis B virus 84
Brucella sp. 81
Salmonella sp. 66
Shigella sp. 56
Hepatitis non-A, non-B 28
Cryptosporidium sp. 27
Total 1074
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LAIs in US, 1979-1999Category Overt LAI Subclinical
LAITotal LAI No. of
deaths
Bacteria 507 40 547 5(4a)
Rickettsia 185 214 399 1
Viruses 523 406 929 1(1a)
Parasites 47 3 50 11
Fungi 5 0 5 0
Total 1267 663 1930 22
(a) Fetus aborted as a consequence of LAI
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Most Common Routes of Exposure
Inhalation of aerosols generated by accident or by work practices
Percutaneous inoculation Contact between mucous membranes and
contaminated material Ingestion
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Accidents Use of needles and syringes Spills and sprays Injuries with broken glass or other sharp
objects Aspiration through pipettes Bites or scratches of animals or
ectoparasites
30Injection Preparation Table,1998
No re-use of disposable injection equipment
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Example: recaped syringes
2003
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Example: mouth pipetting
2005
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Aerosols and droplets are main sources of contamination
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Episodes of Single-source, Multiple Laboratory Infections
Disease Probable Source of Infection
Maximum Distance From Source
Number Persons Infected
Brucellosis Centrifugation Basement To 3rd floor
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Coccidioidomycosis Culture transfersolid media
2 Building floors 13
Coxsackle Virus infection
Spilled tube of infected mouse tissue on floor
5 feet (estimated) 2
Murine Typhus Intranasal inoculation of mice
6 feet (estimated) 6
Tularemia 20 petri plates dropped 70 feet 5
Venezuelan encephalitis
9 lyophilized ampoules dropped
4th floor stairs to 3rd or 5th
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Reitman and Wedum, 1956
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Aerosols from laboratory equipment(1010/ml culture – 10 min. use)
Blender, opened at once 106
Sonicator, with bubbling 106
Pipetting, vigorous 106
Dropping culture 3 x 105
Splash on centrifuge rotor 105
Drop spill on zonal rotor 2 x 104
Blender, opened at 1 minute 2 x 104
Pipetting, carefully 104
Dimmick et al., 1973
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Aerosols from Animal Cage Cleaning
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Samples performed by the patient Stool Urine SputumExternal contamination of sample vial
Samples performed by professional Pus CSF Nasopharyngeal aspirate or bronchoalveolar lavage Nasopharyngeal swab or throat swab BloodRisk at sampling ( needle, contact with skin specially in case of
wounds). External contamination of sample vial
Contamination risks on biological samples
3. Importance of using appropriate biosafety
equipment
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Using appropriate biosafety equipmentEach and every Biological sample is potentially infectious
It is a risk for every person who will handle it
During samplingDuring transport
At the openingDuring handling in the laboratory
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Personal Protective Equipments
Gloves Coat Mask Glasses Screen Biosafety cabinets ?
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Precautions during sample collection
Protect collector, colleague, staff... wear gloves, coat, (mask, glasses)
VHF: double gloves, filter-masks, boots
dispose needles in special containers, without re-capping, disinfection (sodium hypochlorite 2.5%), incineration
clean working surfaces (hypochlorite) decontaminate material (hypochlorite
10%)
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During sample collection Consider each sample as infectious
Clean outer tube with 10% diluted household bleach (chlorine)
Wear gloves (two layers) Wear lab coat, mask and protective glasses, Use evacuated tubes (vacutainers) for blood sampling
Organize and disinfect bench space with Chlorine 10% followed by wash with 70% alcohol Clean spills with Chlorine 10% Decontamination of equipment by Soaking in
Chlorine10%
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Preparation and handling of samples Opening under Biosafety Cabinet Use paper or cotton soaked with ethanol 70% to
handle the cap when opening the tube (this will stop contamination by liquid droplets)
Centrifuge at low speed before opening if liquid is spread all over the tube: check the centrifuge before, balance the tube and use capped buckets.
Never use glass pipets for handling the specimens. No mouth pipetting allowed If viruses are involved, freeze the aliquots as soon
and as cold as possible. For bacteriology keep samples at room temperature
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In all diagnostic and health-care laboratories;
Sharp containers
Specific waste disposal
Never manipulate directly broken tubes
Manipulation of sharps and needles
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Waste disposal
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Good Biosafety Practices
TargetSource
Technicians Samples Support staff
Environment
Sampling
Transport
Opening
Handling in lab
Waste disposal
Three critical steps
4. Four biosafety levels
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Risk Group I
Low individual and community risk
A microorganism unlikely to cause human or animal disease
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Risk Group II
Moderate individual risk, limited community riskA pathogen causing human or animal disease unlikely to be a serious hazard to laboratory staff, the community, livestock or the environmentMay cause serious infection but effective treatment is available and risk of spread is limited
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Group II Examples
Viruses, fungi, parasites, bacteria Legionella Shigella Hepatitis B
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Risk Group III
High individual risk, low community risk
A pathogen producing serious human or animal disease but not readily transmitted to others
Effective treatment available
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Group III examples Viruses, bacteria, fungi
SARS and rabies virus Brucella species Bacillus anthracis Yersinia pestis Multi Drug Resistant Mycobacterium tuberculosis H5N1
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Risk Group IV
High individual and community risk
A pathogen producing serious human or animal disease, readily transmitted from one individual to another.
Effective treatment usually not available
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Group IV Examples
Viruses Ebola Nipah Hendra
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Factors To Consider In Classification
Pathogenicity of the agent Modes of transmission and host range of
organism Local availability of preventive measures Local availability of effective treatment.
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Biosafety levels
Assignment of agent must be based on risk assessment.
Depends on agent and conditions of use Requires professional judgment
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Elements of Containment Primary containment
Protection of personnel and the immediate laboratory environment.
Use of laboratory practices, techniques, safety equipment
Secondary containment Protection of the environment external to
immediate laboratory
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Risk Groups vs. Biosafety Levels (1)
Risk Group Biosafety Level
Laboratory Type
Laboratory Practices
Safety Equipment
1 Basic- Biosafety Level 1
Basic teaching, research
GMT None; open bench work
2 Basic – Biosafety Level 2
Primary health services; diagnostic services, research
GMT plus protective clothing, biohazards sign
Open bench plus BSC for aerosols
BSC= Biological Safety CabinetGMT= Good Microbiological Techniques
* Laboratory Biosafety Manual, 3rd edition, 2005 .
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Risk Groups vs. Biosafety Levels (2)Risk Group 1 Biosafety
LevelLaboratory Type
Laboratory Practices
Safety Equipment
3 Containment- Biosafety Level 3
Special diagnostic services, research
Level 2 + special clothing, access control, directed airflow
BSC and/or other primary devices for all activities
4 Maximum Containment – Biosafety Level 4
Dangerous pathogen units
Level 3 + airlock entry, shower exit, special waste disposal
Class III BSC, or positive pressure suites with class II BSCs, double ended autoclave
BSC= Biological Safety CabinetGMT= Good Microbiological Techniques
* Laboratory Biosafety Manual, 3rd edition, 2005 .
5. Risk assessment
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Risk
Risk of infection is defined as chance of exposure to hazard or exposure to chance of injury
Risk of infection may be quantitative or qualitative
Laboratory Director’s responsibility
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Criteria for risk assessment Pathogenicity (the ability to cause disease) and
disease severity Infectious dose (lower the dose, the higher the
hazard) Transmission (the importance of aerosols) and
agent stability in the environment Volume and concentration (risk increases) Laboratory activity planned, including potential
genetic manipulation Presence of suitable host Prophylaxis (local availability of vaccine or
treatment) and medical surveillance
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If limited information on agent or specimen
Use standard precautions Use basic containment - BSL 2 Follow national/international regulations
for transport Assess medical data Get epidemiological data Determine geographical origin of agent
6. Appropriate biosafety documentation
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Awareness, Training, Vigilance
Awareness – Advise workers of possible exposures, safeguards and responsibilities
Training – Inform workers of hazards of their work, and use of appropriate practices, techniques and procedures
Vigilance – Maintain vigilance to guard against safety procedure compromise or errors.
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Good biosafety
Good laboratory practices Awareness of hazards Knowledge of how laboratory infections
occur Knowledge of procedures and techniques
to reduce hazards
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But also
Biosafety Officer
Biosafety manual
Biosafety procedures
Risk assessment
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Total Quality
Improving all internal and external processes that contribute to the final product/service
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Total Quality Principles
Requires: the existence of a Supervisor responsible Standards Operating Procedures the training of the personnel
aware of potential defaulsaware of and applying procedure
Quality manual specific to different laboratories
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Biosafety
The application of combination of laboratory practices and procedures,
laboratory facilities and safety equipment
when working with potentially infectious microorganisms
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Biosafety Principles Requires
the existence of a Supervisor responsible Standards Operating Procedures the training of the personnel
aware of potential hazardsaware of and applying practices and techniques
Biosafety manual specific to different laboratories
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Biosafety Practices Plan Written procedures on the proper way to
do/remove Personal Protective Equipment Entering/leaving isolation suites
Protocols on the use of Biosafety Cabinets Decontamination protocols
Spills Surface Instruments
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Emergency procedures Develop an emergency plan
Cuts, accidental punctures Ingestion of chemicals/hazardous materials Spills Broken/leaking containers Fire, and other natural disaster Facility emergencies
Power outages
Ventilation and Filters failure
Flood Threats
Appropriate documentation on disinfectants Emergency Contact Information Appropriate and complete medical records should be kept Write an Incident Report
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Incident Report Form Relevant information
Date and person reporting the incident Description of the incident Assessment of the problem Corrective Action Reviewed by the Supervisor/Lab Director
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Biosafety in Microbiological and Biomedical Laboratories
BMBL 4th Edition CDC/NIH Canadian Laboratory
Biosafety Guidelines WHO Biosafety
Guidelines ISO 15 190 Documentation on
disinfectants
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Summary
The laboratory must have a restricted entrance The staff must be aware of the risks The staff must be trained to biosafety The staff must use proper biosafety protective
equipment There must be a supervisor There must be standard operating procedures All documents concerning biosafety must be
available
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Summary
All diagnostic and health-care laboratories must be designed and organised for Biosafety level 2 or above.
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Problem Scenario
Your laboratory is specialized in virology. There has been an important epidemic of H5N1 in your country and the Ministry of Health has asked you to be the referent laboratory for this pathology. What must you do to ensure biosafety in your laboratory?
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