ࡱ> kmhiji y^bjbj :<{b{b[U.(.(.(.(.(B(B(B(8z()B(3++:,,,,,,`-^-$-3333333$68\43.(-`-`---43.(.(,,,,4I3X6X6X6-.(,,.(,,3X6-3X6X6. H,,,i.R 2_303 '92'9,,'9.(-tX6---43435---3----'9---------B &: Appendix 4 Biosafety SOP Author: Penny StevensDocument Number:Fac10-13-App 4Effective (or Post) Date:17-Feb-09Review HistoryDate of last review:13 Feb 2020Reviewed by:Heidi HanesSMILE Comments: This document is provided as an example only. It must be revised to accurately reflect your labs specific processes and/or specific protocol requirements. Users are encouraged to ensure compliance with local laws and study protocol policies when considering the application of this document. If you have any questions contact SMILE. Appendix 4 Biosafety SOP Author(s), Name & TitlePenny S. Stevens MBS, MT (ASCP), CLS (NCA)Document NumberEffective DateInternational QA/QC CoordinatorFac10-13-SOP Appendix 417 Feb 2009SMILE Comments: This document is provided as an example only. It must be revised to accurately reflect your labs specific processes and/or specific protocol requirements. Users are directed to countercheck facts when considering their use in other applications. If you have any questions contact SMILE. Approved By Name, TitleSignatureDate SOP Annual ReviewName, TitleSignatureDate Revision HistoryVersion # [0.0]Revision Date [dd/mm/yy]Description (notes)2.017 Feb 2009Reformatted to meet SMILE Resource Template format requirements. Distributed Copies toName (or location)# of copiesName (or location)# of copies I acknowledge that I have read, understand and agree to follow this SOP. Name (print)SignatureDate APPENDIX 4 PRINCIPLES OF BIOSAFETY DEFINITIONS BSC - Biological Safety Cabinet BSL - Biological Safety Level PPE Personal Protective Equipment UV Ultraviolet Light TABLE OF CONTENTS Containment Primary Containment Secondary Containment Biosafety Levels Table of Biological Safety Levels Biological Safety Cabinet Containment The term "containment" is used in describing methods for managing infectious agents in the laboratory environment where they are being handled or maintained. The purpose of containment is to reduce exposure of laboratory workers, other persons, and the outside environment to potentially hazardous agents. The elements of containment include laboratory practices and techniques, safety equipment, and facility design. Primary containment, the protection of personnel and the immediate laboratory environment from exposure to infectious agents, is provided by good technique and the use of appropriate safety equipment. Secondary containment, the protection of external laboratory environment from exposure to infectious materials, is provided by a combination of facility design and operational practices. primary containment The most important element of primary containment is strict adherence to standard biohazard safety practices and techniques. Persons working with infectious agents or materials must be aware of potential hazards and be trained and proficient in the practices and techniques required for handling such material safely. The supervisor is responsible for providing or arranging for appropriate training of personnel. Additional measures may be necessary when standard laboratory practices are not sufficient to control the hazard associated with a particular agent or laboratory procedure. The selection of additional safety practices is the responsibility of the laboratory supervisor and must be commensurate with the inherent risk associated with the agent or procedure. Each laboratory must develop or adopt a safety manual, which identifies the hazards that may be encountered and specifies practices designed to minimize or eliminate risks. Personnel should be advised of special hazards and should be required to read and follow the required practices and procedures. In the Microbiology Laboratory, activities must be supervised by a microbiologist who is trained and knowledgeable in appropriate laboratory techniques, safety procedures and associated risks. Laboratory personnel safety practices and techniques must be supplemented by appropriate facility design and engineering features, safety equipment, and management practices. Biosafety Levels An important element of secondary containment is the use of Biosafety Levels (BSLs). These guidelines specify four BSLs with the potential safety hazards posed by the infectious agents for which the laboratory is responsible. The levels are determined by of a combination of laboratory practices and techniques, potential hazard posed by the infectious agents, degrees of protection provided to personnel, safety equipment, and the laboratory facilities. The object of these guidelines is to inform the laboratory staff of the safety practices required when handling potentially hazardous organisms and biological materials. Each laboratory worker is responsible for his/her own safety, the safety of his/her fellow worker and training in the safety methods used in the laboratory. Remember: the most expensive equipment is not a substitute for careful technique. Biosafety Level 1. (BSL-1) - Safety equipment and facilities must be appropriate for laboratory personnel with specific training in the procedures conducted in the laboratory. The personnel should be supervised by a scientist with general training in microbiology or a related science. Work in these areas will generally be conducted on open bench tops using standard microbiological practices. Special containment equipment or facility design is neither required nor generally used. Standard Microbiological Practices a. Access to laboratory should be limited or restricted at the discretion of lab director when work or experiments on cultures and specimens are in progress. b. A biohazard sign should be posted at the entrance to the laboratory. The sign should include the name of the agents in use and the names and phone numbers of the lab contacts. c. Lab coats, gowns or uniforms should be worn to prevent contamination or soiling of street clothes and should remain in the laboratory unless decontaminated. d. Gloves (non- latex) should be worn. e. Protective eyewear must be worn for procedures in which splashes of microorganisms or other hazardous materials are anticipated. f. Procedures are performed to minimize splashes or aerosols. g. Staff must wash their hands after handling viable materials, after removing gloves, and before leaving lab. h. Work surfaces must be decontaminated at least once a day and after any spill of viable material. i.All cultures, stocks, and other regulated wasted must be decontaminated before disposal by an approved decontamination method such as autoclavingj. Refer to the General Lab Safety Policy (Fac1.0-13) for additional safety requirements.  Facilities a.Lab doors have access control.b.Each lab has a sink for hand washing.c.Lab is easily cleaned. No carpet or rugs in lab area.d.Bench tops are impervious to water.e.Spaces between benches, cabinets, and equipment are accessible for cleaning. Biosafety Level 2. (BSL-2) - This level should be adopted when work involves agents of moderate potential hazard to personnel and environment. Standard Microbiological Practices: a.All BSL-1 requirements.b.Work surfaces are decontaminated with material specifically effective against the agent of concern. Special Practices: a.Biohazard sign must be posted on entrance to lab when etiologic agents in use. The sign must include names and telephone numbers of lab contacts, personnel protective equipment required in lab, agents or microbes in use and biosafety level of lab.b.Lab personnel must receive appropriate immunizations or tests for the specific agents handled. When appropriate a base line serum sample is collected and stored.c.Biosafety procedures are incorporated into standard operating procedures. Personnel are advised of special hazards. d.Lab director ensures the lab personnel receive appropriate training on potential hazards associated with work involved and precautions to prevent exposure and evacuation procedures. Personnel receive annual updates or training as necessary for policy and procedure changes. e.Use a high degree of caution with any contaminated sharp items, including needles and syringes, slides, pipettes, capillary tubes, and scalpels. Substitute plastic for glassware whenever possible. f.Cultures, tissues, body fluid specimens, or potentially infectious wastes are placed in a container with a cover that prevents leakage during collection, handling, processing, storage and transport. Safety Equipment and Facilities They should be applicable to indigenous moderate-risk agents present in the community and associated with human disease of varying severity. Organisms and activities with low aerosol potential can be conducted on the open bench using good microbiological techniques i.e., hepatitis agents, salmonellae, and Toxoplasma spp. Primary barriers include: Biological safety cabinets, splash shields, face protection, protective lab coats, gowns and gloves. Secondary barriers include: Hand washing and waste decontamination facilities to reduce potential environmental contamination. Eyewash station is readily available. Furniture is covered with non-fabric material that can be decontaminated. Lockable doors are provided for restricted agents. Examples of high-risk steps in the laboratory would include: a.Specimen Collection (e.g. needle sticks)b.Specimen Processing (e.g. spills in transit, aerosols from improper centrifugation, removal of stoppers, decanting of serum or plasma with external contamination of containers and/or work surfaces)c.Specimen Analysisd.Disposal of Specimen (e.g. failure to separate specimen containers from non-infectious laboratory waste)e.Procedures with high aerosol potential may predictably and significantly increase the risk of exposure of personnel to infectious aerosols and must be conducted in primary containment equipment or devices.  Biosafety Level 3. (BSL-3) - Applicable to work with indigenous or exotic agents, which may cause serious and potentially lethal infections or disease as a result of exposure by inhalation i.e., Mycobacterium tuberculosis, St. Louis encephalitis virus, and Coxiella burnetii Standard Microbiological Practices all BSL-1 and BSL- 2 restrictions apply. Special Practices a.Laboratory doors are kept closed when work is in progress.b.The laboratory director controls access and restriction to the lab.c.Biosafety manual must be specific to the laboratory and prepared or adopted by the lab director and biosafety precautions are incorporated in the procedures. d.All manipulations involving infectious material are conducted in biological safety cabinets. Clean up is facilitated by using plastic backed paper toweling on non-perforated work surfaces within biological safety cabinets.e.Equipment must be decontaminated before removal from the facility for repair or maintenance or packaging for transport.f.All spills and exposures are reported to the laboratory director. Appropriate medical evaluations, surveillance, and treatment are provided and records maintained by management. Safety Equipment (Primary barriers) include: a.Biological safety cabinets (BSC) or other enclosed equipment must be used for ALL laboratory manipulations. No culture work should be done on open benches.b.Protective clothing such as solid front or wrap-around gowns, scrub suits, or overalls must be worn by workers in the lab. Along with all barriers listed under BSL-1 and BSL-2.c. Laboratory clothing that protects street clothing (i.e., solid front or wrap-around gowns, scrub suits, coveralls, etc.) must be worn in the laboratory. FRONT-BUTTON LABORATORY COATS ARE UNSUITABLE. Laboratory clothing must not to be worn outside of the laboratory and must be decontaminated before laundered. Laboratory Facilities (Secondary barriers) include: a.The lab is separated from areas with unrestricted traffic. Access to the laboratory is controlled. Laboratory doors are kept closed when cultures are being processed or identified. Access must be through a set of self-closing double doors.b.A ducted exhaust air ventilation system must be provided and a specialized ventilation system that creates a directional airflow which draws air into the laboratory from clean areas toward contaminated areas. This minimizes the release of infectious aerosols from the laboratory to clean areas.c.Biosafety cabinets are required and must be located away from doors, ventilation systems, and from heavily traveled lab areas. d.All windows must be closed and sealed. The interior surfaces of walls, floors, and ceilings of areas where BSL- 3 agents are handled must be constructed for easy cleaning and decontamination. Seams, if present, should be sealed. All surface areas should be impermeable to liquids and resistant to damage from the chemicals and disinfectants normally used in the laboratory.e.The laboratory supervisor will assure that only persons who have been advised of the potential biohazard, meet any of the specific entry requirements (e.g. immunization and baseline serum), and comply with all entry and exit procedures are permitted to enter the laboratory.f. When infectious materials are present in the laboratory, a hazard warning sign, incorporating the universal biohazard symbol, is posted on all laboratory access doors and on other items (i.e., equipment, containers, materials, etc.) as appropriate to indicate the presence of viable infectious agents. The hazard warning sign should identify the agent, list the name of the laboratory supervisor and another responsible person(s), and indicate any special conditions of entry into the area (immunizations, respirators, etc.). Primary hazards to personnel working with these agents include auto- inoculation, ingestion, and exposure to infectious aerosols. Examples of high-risk steps in the laboratory are the same as BSL-1 & 2. Biosafety Level 4. (BSL-4) - Applicable to working with dangerous and exotic agents, which pose a high individual risk of life-threatening disease. All manipulations of potentially infectious diagnostic materials, isolates, and naturally or experimentally infected animals, pose a high risk of exposure and infection to laboratory personnel. Lassa fever and Ebola viruses are examples of BSL-4 microorganisms. This level is not applicable to the diagnostic laboratories. BIOSAFETY LEVEL TABLE BSLAgentsPracticesSafety Equipment (Primary Barriers)Facilities (Secondary Barriers)1Not known to consistently cause disease in healthy adultsStandard Microbiological practicesLab coats, gowns or uniforms Gloves, protective eyewear where potential splashes anticipatedOpen bench top & sink required2Associated with human disease. Hazards are percutaneous. Injury, ingestion, & mucous membrane exposureBSL-1 plus limited access Biohazard warning signs. Sharps precautions, & biosafety manualPrimary barriers: BSC or physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials. PPEs: lab coats, gloves, & face protection as neededBSL-1 plus: Autoclave available3Indigenous or exotic agents with potential for aerosol transmission. Disease may have serious or lethal consequences.BSL-2 plus: controlled access, decontamination of all waste, decontamination of lab clothing before laundering, & baseline serum testsPrimary barriers: BSC or other physical containment devices used for all open manipulations of agents. Standard PPE plus: additional protective lab clothing and respiratory protection as neededBSL1 &2 plus: Physical separation from access corridors ,Self-closing double door access, exhausted air not re-circulated and negative airflow lab4Not applicable to the diagnostic laboratory BIOLOGICAL SAFETY CABINET - BSCs are designed to provide personnel, environment and product protection when appropriate practices and procedures are followed. Three kinds of biological safety cabinets, designated as Class I, II and III have been developed to meet varying clinical needs. Class I - has negative pressure with minimum face velocity of 75 linear feet per minute (Lfmp) and all of the air from the cabinet is exhausted through a HEPA filter either into the laboratory or to the outside. Class I BSCs are no longer being manufactured on a regular basis and many have been replaced by Class II BSCs. Class I BSCs may be used for centrifuges, harvesting equipment or blenders but do not provide a microbe free work environment. Class II Personnel protection is provided with the air flow being drawn around the operator inward with a face velocity of 75 - 100 Lfpm, HEPA - filtered vertical laminar airflow provide product protection by minimizing cross-contamination along the work surface of the cabinet, and HEPA filter exhaust air for environmental protection. All Class II cabinets are designed for work with microorganisms assigned biosafety levels 1, 2, and 3. They provide a microbe free work environment. They are not to be used with volatile or toxic chemicals. An example of the Class II vertical laminar-flow biological cabinet (type A) is an open-fronted, ventilated cabinet with an average inward face velocity at the work opening of at least 75 feet per minute. This cabinet provides a HEPA-filtered, recirculated mass airflow within the work space. The exhaust air from the cabinet is also filtered by HEPA filters. Design, construction, and performance standards for Class II cabinets have been developed by and are available from the National Sanitation Foundation, Ann Arbor, Michigan. Class III - is totally enclosed, ventilated cabinet of gas-tight construction and has the highest degree of personnel and environmental protection from infectious aerosols, as well as protection of research materials from microbiological contamination. Used mostly for work with hazardous agents that requires Biosafety levels 4 containment. All work is done through attached rubber gloves and the cabinet is operated under negative pressure. Supply air is HEPA filtered, and cabinet exhaust air is filtered by two HEPA filters in series. Class III must be connected to double-doored auto claves and chemical dump tanks to sterilize or disinfect all materials exiting the cabinet. BSC effectiveness is a function of directional air flow (inward and downward), through a "high efficiency particulate air" (HEPA) filter. Rapid movement can disrupt the airflow and reduce effectiveness i.e., rapidly moving your arms in and out of the BSC and people walking rapidly behind you. For best results, Class I and II BSCs should be located away from traffic patterns, doors, ventilation systems, and air handling devices. BSC Operation: Do NOT place objects on or over front air intake grille. Do NOT block rear exhaust grille. Arrange materials to segregate contaminated and clean items. Work should be performed at least six (6) inches back from front grille. Inside the BSC, always use horizontal pipette discard pans, containing appropriate disinfectant. Clean up all spills immediately. Wait 5 minutes before resuming work. BSC Maintenance: Cabinets should be decontaminated at least once per day after completion of work processes. UV Lights should be maintained as indicated in Fac1.0-13 Appendix 5-Electrical & Mechanical Safety. Cabinets must be certified at least annually to ensure that filters are functioning properly and that airflow rates meet required specifications. Resources NCCLS. Clinical Laboratory Safety; Approved GuidelineSecond Edition. NCCLS document GP17-A2 [ISBN 1-56238-530-5]. NCCLS, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2004. Clinical and Laboratory Standards Institute. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline-Third Edition. CLSI document M29-A3 [ISBN 1-56238-567-4]. Clinical and Laboratory Standards Institue, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087-1898 USA, 2005. College of American Pathologists (CAP) 2006. Commission on Laboratory Accreditation, Laboratory Accreditation Program; Laboratory General Checklist Revised 9/27/2007. College of American Pathologists (CAP) 2006. Commission on Laboratory Accreditation, Laboratory Accreditation Program; Microbiology Checklist Revised 9/27/2007. CDC-NIH U.S. Department of Health and Human Services Primary Containment for Biohazards: Selection, Installation and Use of Biological Safety Cabinets, Sept 2000, 2nd Edition. ACGIH (American Conference of Governmental Industrial Hygienists) Threshold Limit Values. 1994-1995. Cincinnati, OH. McKinney, Robert, Richard Jonathan. CDC/NHI Department of Health and Human Services, Biosafety in Microbiological and Biomedical Laboratories 4th Ed. May 1999. U.S. Government Printing Office. H.H.S. Publication No. (CDC) 93-8395. Occupational exposure to hazardous chemicals in laboratories, OSHA laboratory standards 29CFR1910.1450 Infection Control: The Johns Hopkins Hospital Interdisciplinary Clinical Practice Manual (ICPM) Infection Control Policy 1998, Osler 4, The Johns Hopkins Institutions Office of Health, Safety and Environmental, Johns Hopkins Safety Manual. 2001, 2024 E. Monument St. Telephone 955-5918 CDC-NIH U.S. Department of Health and Human Services Biosafety in Microbiological and Biomedical Laboratories, May 1999, 4th Edition. (HHS Publication No. (CDC) 93-8395).     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