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Laboratory BiosafetyLaboratory Biosafety

WSLH TeleconferenceJune 27, 2007

Peter A. Shult, Ph.D.Director, Communicable Disease Division

and Emergency Laboratory ResponseWisconsin State Laboratory of Hygiene

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Laboratory BiosafetyLaboratory Biosafety

• Historical perspective• Principles of biosafety

• Elements of containment including the Biological Safety Cabinet (BSC)

• Biosafety levels• Biosafety risk assessment• Biosafety beyond the laboratory walls• Biosecurity

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Laboratory BiosafetyKey Resources (I)

• Biosafety in Microbiological and Biomedical Biosafety in Microbiological and Biomedical Laboratories (BMBL),5Laboratories (BMBL),5thth Ed. Ed.

U.S. Department of Health and Human Services http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm

http://www.slh.wisc.edu/wps/wcm/connect/extranet/comdis/

• Laboratory Biosafety Manual, 3rd Ed. World Health Organization, 2004

http://www.who.int.csr/resources/publications/biosafety/

WHO_CDS_CSR_LYO_2004_11/en/

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Laboratory BiosafetyKey Resources (II)

• Primary Containment for Biohazards: Selection, Installation and Use of Biological Safety Cabinets, 2nd Edition

U.S. Department of Health and Human Services Public Health Service Centers for Disease Control and Prevention and National Institutes of Health September 2000 http://www.cdc.gov/od/ohs/biosfty/bsc/bsc.htm

• Control of Communicable Disease Manual, 18th Edition David L. Heyman, MD, Editor APHA

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Laboratory BiosafetyKey Resources(III)

• Public Health Guidance for Community-Level Preparedness and Response to Severe Acute Respiratory Syndrome (SARS),Version 2.3; July 20 2004

http://www.cdc.gov/ncidod sars/guidance

• Biological Safety: Principles and Practices, 4th Ed ASM Press, 2006• Laboratory Security and Emergency Response Guidance for Labs Working with Select Agents MMWR, December 6, 2002; Vol.51/ No. RR-19 http://www.cdc.gov/mmwr/PDF/RR/RR5119.pdf

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Laboratory BiosafetyHistorical perspective (I)

• Landmark studies by Pike and Sulkin• Questionnaire assessment• Between 1930 – 1978, 4,079 LAIs with 168 deaths• Most common causative agents of overt infection include: 1. Brucella spp. 6. M.tuberculosis 2. C. burnetti 7. B. dermatidis 3. HBV 8. VEE 4. S. typhi 9. C.psittaci 5. F. tularensis 10. C.immitis• No specific accident or exposure event in > 80%

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Laboratory BiosafetyLaboratory BiosafetyHistorical perspective (II)Historical perspective (II)

• Followup worldwide literature search, 1979-2004• 1, 141 overt infections, 24 deaths• Most common causative agents of overt infection include: 1. M. tuberculosis 6. HBV 2. Arboviruses 7. Shigella spp. 3. C. burnetti 8. Salmonella spp 4. Hantavirus 9. HCV 5. Brucella spp. 10. N. meningitidis• Many asymptomatic infections• Many newer agents, e.g. SARS-CoV, Cryptosporidium, etc.• No specific exposure event in most cases

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Laboratory BiosafetyHistorical perspective (III)

What types of laboratories involved ? % of LAIs according to laboratory type 1930-1975 1979-2004 Clinical 17% 46%Research 59% 50%Production 3% 3%Teaching 3% 1%Unknown 18% < 1%• Reasons for increase in LAIs in clinical labs?

• Better surveillance and reporting• Absence of biosafety containment equipment• Failure to use adequate containment procedures early in

diagnostic process

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Laboratory BiosafetyLaboratory BiosafetyHistorical perspective (IV)Historical perspective (IV)

What were the predominant means of exposure?• As mentioned before no specific exposure event

identified in most cases• Those identified included:

• Inhalation of aerosols generated by work practices or procedures or spills

• Percutaneous inoculation• Contamination of mucous membranes• Ingestion

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Laboratory BiosafetyPrinciples of Biosafety

• The objective of biosafety is the containment of potentially harmful biological agents

• The purpose of containment is to reduce/eliminate exposure of lab workers, other persons and outside environment to biohazardous agents

• Key elements of containment include:• Laboratory practice and technique• Safety equipment (primary barriers and PPE)• Facility design and construction (secondary

barriers)• Risk Assessment of the work to be done with a specific

agent or under specific circumstances determines the appropriate combination of these elements to employ

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Laboratory BiosafetyElements of Containment

Laboratory Practice and Technique• Arguably the most important element of containment• Awareness of potential hazards and training and experience

are critical• Applies to pre-analytical, analytical and post-anaytical

processes• What about non-traditional testing sites and personnel?

• Behavioral factors• Need for clear focus on work – all ages• “Creative innovation” and risk taking

• Suggested age relation• Lose the bad habits: sniffing, “hot looping”, etc.• Working in absence of other containment elements

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Laboratory BiosafetyElements of Containment

Safety Equipment (Primary barriers and PPE)• Available for each possible route of exposure

• Aerosol: BSCs, covered centrifuge carriers, loop incinerators or disposable loops, PPE (respirators, PAPRs)

• Percutaneous: sharps disposal; retractable needles• Mucous membrane contact: goggles or safety glasses,

face shields, gloves• Ingestion: automatic pipetting devices

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Laboratory BiosafetyElements of Containment

The Biological Safety Cabinet (I) Arguably the single most important piece of safety

equipment in the laboratory!• Information Resources• Importance of the Biological Safety Cabinet

• The principal device used to provide containment of infectious splashes or aerosols generated by many microbiological procedures

• Provides protection to the operator, the laboratory environment and work materials

• Which type is for you?

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Laboratory BiosafetyElements of Containment

The Biological Safety Cabinet (II)

• Follow proper BSC work practices and procedures• BMBL5, Appendix A

• They don’t work at all if you don’t use them!

• Need for a lab-specific algorithm for BSC use• A POLICY ISSUE• Tie into risk assessment

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Laboratory BiosafetyElement of Containment

Facility Design and Construction (Secondary Barriers)• Contributes to laboratorian safety, however, primary

role is to protect persons outside of the lab and persons in the community from agents that might be accidentally released.

• Recommended secondary barriers depend on the risk of transmission of specific agents• BSL-1/BSL-2 vs. BSL-3

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Laboratory BiosafetyBiosafety Levels

• 4 biosafety levels • Consist of combinations of lab practices and techniques,

safety equipment and lab facilities• Purpose: To categorize risk associated with infectious agent and

define the appropriate safety practices, equipment and facilities for handling the agent safely

• Appropriate BSL determined by:• Microbiological agent Risk Group• Mode of transmission• Procedural protocols• Experience of staff• Likelihood of aerosol generation• Work involves use of amplified agent• Other?

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W.H.O. Agent Risk Group Classification

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BSL Agents Microbiology Practices

Safety Equipment (Primary Barriers)

Facilities (Secondary Barriers)

1

Not known to consistently cause disease in healthy adults

Standard Microbiological Practices

None required Open bench top sink required

2

Associated with human disease, hazard equals percutaneous injury, ingestion, mucous membrane exposure

BSL-1 plus: Limited access, biohazard warning signs, “sharps” precautions, biosafety manual defining waste decontamination & medical surveillance policies

Class I or II BSCs or other physical containment for manipulations of agents that cause splashes or aerosols of infectious materials, PPE: lab coats, gloves, face protection as needed

BSL-1 plus: Autoclave

3

Indigenous or exotic agents with potential aerosol transmission; may have serious or lethal consequences

BSL-2 plus: Controlled access; Decontamination of all waste & lab clothing before laundering; Baseline serum

Class I or II BCSs or other physical containment for all manipulations; PPE: protective clothing; gloves; respiratory protection as needed

BSL-2 plus: Physical separation from corridors; Self-closing, double-door access; Exhausted air not recirculated; Negative airflow into laboratory

4

Dangerous/exotic agents with high risk of life-threatening disease, aerosol-transmitted infections; or related agents with unknown risk of transmission

BSL-3 practices plus: Clothing change before entering, shower on exit, all material decontaminated on exit

All procedures in Class III BSCs or Class I or II BSCs in combination with full-body, air-supplied, positive pressure personnel suit

BSL-3 plus: Separate building or isolated zone, dedicated supply, exhaust, vacuum, & decon systems; other requirements.

Recommended Biosafety Levels for Infectious Agents“Biosafety in Microbiological and Biomedical Laboratories”,5th Ed

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Laboratory Biosafety

Risk Assessment

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Handling Unusual Test Requests in the Clinical Lab

What is meant by “unusual test request”?

• “Novel” agents, high public health impact• SARS, Monkeypox, Avian influenza, smallpox

• Agents of particular public health importance• Mumps, measles, rubella, hantavirus, etc.

• “Novel” agents, impact uncertain• hMNV, Coronaviruses (non-SARS), Bocavirus, HPV

• Rare or exotic agents• B virus, HFVs, chikungunya, dengue, etc.

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Handling Unusual Test RequestsHandling Unusual Test Requests Considering the request

Assessments needed• Type of request• Diagnostic capability

• Does it exist in lab? Should it be used?• Can it/should it be developed?

• Biosafety considerations• Expertise and Experience• Regulatory restrictions on testing

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Laboratory BiosafetyRisk Assessment(I)

• Whose responsibility?• Technically, the lab director • Practically, the bench microbiologist

• Primary factors to consider:• Agent hazards• Laboratory procedures planned

• Potential for aerosol generation• Consider facility, equipment needed; appropriate PPE

• Capability of the staff • Training, technical proficiency, good habits

• Known vs. unknown agent risk• For known or suspected agent, consult BMBL agent summary

statements, other references• For unknown agent…

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Laboratory BiosafetyRisk Assessment(II)

Risk Assessment for Unknown Agents• Reason for the need

• The Age of Emerging Diseases • SARS, avian influenza, influenza A (H2N2)

• Key element of the assessment• In addition to above, patient information is critical

• Evaluate completeness of patient information to assess risk of specimen testing

• When should it be undertaken? • During emergency response (BT or EID) vs. routinely?

• Need new biosafety model in the laboratory!• Standard (“Universal”) precautions for blood and

fluids• Enhanced precautions for respiratory (and other?)

specimens needed???

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Laboratory BiosafetyRisk Assessment(III)

Other Considerations in the Clinical Laboratory• Strict BSL-2 practices and procedures should be the minimum

standard• Biosafety cabinets (BSCs) are a must!

• But the reality is…• Needs to be an organizational priority• At minimum, develop algorithm for their use based on

risk• What about a possible BSL3 agent but no BSL3 lab? “Mix &match” PPE and good practice with equipment and

facility based on risk assessment → BSL 2+• Be aware of the potential for exposure to a BSL4 agent• Look for “just in time” guidance, e.g. SARS

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Laboratory BiosafetyBiosafety Beyond the Laboratory Walls

• Specimen collection sites• Specimen transport: route and packaging• Specimen labeling and requisition – “A heads up”• Close communication with ICP and clinicians• Don’t forget your rapid test sites!

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Locations of Sentinel Laboratories and Rapid Test Sites in Wisconsin-2005

Sentinel Laboratories

Douglas Bayfield

Ashland

SawyerWashburnBurnett

Polk Barron Rusk

Price

Iron

Vilas

Oneida

Lincoln

TaylorChippewaSt. Croix

Pierce

Dunn

Pepin

Eau Claire

BuffaloTrempealeau

Jackson

Monroe

Clark Marathon

Wood Portage

Juneau Adams

Sauk

LaCrosse

Vernon

Crawford

Richland

Grant

LaFayetteGreen

Rock

Dane

Iowa

Columbia Dodge

Jefferson Waukesha

Walworth

Kenosha

Racine

Ozaukee

SheboyganFond du Lac

Green Lake

Marquette

Waushara Winnebago

Calumet

Manitowoc

Kewaunee

Brown

OutagamieWaupaca

Shawano

Menominee

OcontoLanglade

Forest

Marinette

Florence

Door

Milwaukee

Washington

Rapid Test Sites

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Laboratory BiosafetyAddressing Rapid Test Site Biosafety Needs

• Obvious need for basic biosafety training• Role for WSLH and clinical labs to participate in training

• Strategies to enhance biosafety & reduce potential exposure• Collect & communicate patient travel history and risk factors

to testing staff• In most cases, no BSC. Therefore:

• Techniques to minimize aerosol production• Consider use of personal protective equipment (PPE)

during test performance• Consider use of physical barriers for test performance

(e.g., bench shields)• Sequester/isolate testing area

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Biosecurity

• What is biosecurity?

• Select Agent Regulations

• Elements of a facility security plan

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BiosecurityWhat is biosecurity?• Protection of high-consequence microbial agents and

toxins, or critical relevant information, against theft or diversion by those who intend to pursue intentional misuse• A concern in light of recent terrorism events

• Relationship to biosafety• Its all about risk assessment and containment!

• Need for general biosecurity planning???• No current federal requirement for such a plan• Excellent review in Section VI of BMBL 5th Ed.

• Enhanced emphasis under Select Agent regulations• Specific requirement for a facility security plan

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Biosecurity Select Agent Regulations

What is the Select Agent Regulationand who is affected?

Establishes a listing of agents thought topose a threat to public safety

Requires entities that possess any ofthese agents to follow the guidelineswithin this regulation

Requires a Facility Security Plan

http://www.cdc.gov/od/sap

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BiosecuritySelect Agent Regulation• Clinical labs likely to be exempt unless they possess S.A.’s• What are diagnostic/clinical labs required to do if they

encounter* a select agent?

• Notification• Contact WSLH • Contact CDC by phone

• Select agent handling protocol• Within 7 days of identification:

• Transfer to registered entity• Destroy---autoclave, incinerate

• Documentation• APHIS/CDC form 4; maintain copy for 3 years• APHIS/CDC form 2 if transferred*

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Biosecurity

Elements of a Facility Security Plan• Required under Select Agent Regulations• Pragmatic applications apart from select agents• Conduct risk assessment as precursor to security plan• Element of plan include:

• Physical security• Data and IT security• Personnel security assessment policy• Controlled access to areas containing select agents• Select Agent accountability including receipt and transfer• Emergency response plan• Incident reporting system

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Laboratory BiosafetyLaboratory Biosafety

Any Questions?Any Questions?