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Njmrc Biosafety Manual UNIVERSITY of DENVER BIOSAFETY MANUAL 2 Table of Contents Introduction pg.3 1. Biosafety Guidelines and Standards pg.4 1.1 CDC/NIH Biosafety in the Microbiological and Biomedical pg. 4 Laboratories pg.4 OSHA Occupational Exposure to Bloodborne Pathogens 1.2 Standard Title 29, CFR Part 1910.1030 1.3 NIH Guidelines for Research Involving Recombinant DNA pg. 5 Materials 2. Biohazard Materials pg. 12 3. Biosafety Levels and Containment pg. 22 3.1 BSL Definitions pg. 22 3.2 Biosafety Cabinets Pg. 31 4. Laboratory Practices and Techniques pg. 39 5. Personal Protective Equipment pg. 43 6. Packaging/Shipping Infectious Materials and pg. 48 Specimens 7. Select Agents pg. 49 8. Decontamination and Disinfection pg. 52 8.1 Chemical disinfection pg. 52 8.2 Steam Sterilization Pg. 56 9. Disposal of Infectious Waste pg. 59 10. DU Waste Guidelines pg. 61 Appendix 1 Risk Groups pg. 67 Appendix 2 Protection of Vacuum Lines pg. 68 Appendix 3 Placement of Materials in BSC pg. 69 Appendix 4 Biohazard Symbol pg. 70 Appendix 5 Reference Materials pg. 71 Version 3.0 2020 3 Introduction DU BIOSAFETY MANUAL It is the goal of the University of Denver (DU) to provide a safe environment for students, faculty, staff, and the environment. The guidelines and recommendations indicated in this manual will help ensure the safety of personnel and the workplace as it relates to biological materials. I. Scope These policies are for all investigators and staff who conduct, or are engaged in biological or biomedical studies at DU. A. These policies are intended for work that involves the use of recombinant DNA, human blood/tissues, animal blood/tissues, and other potentially infectious agents. B. The policies will address compliance with the NIH Guidelines for Research Involving recombinant DNA, OSHA Bloodborne Pathogens Standard, the CDC-NIH Biosafety in Microbiological and Biomedical Laboratories, and the OSHA Laboratory Standard and the Hazard Communication Standard, as applicable. C. Good laboratory practices and standard precautions should be practiced in any laboratory where biomedical research is being conducted. This document is written as a guideline for work in the research. It addresses engineering controls, work practices and personal protective equipment that should be implemented to minimize the exposure to infectious agents or other potentially infectious material, recombinant DNA, or material and/or its toxin that is known or suspected to cause human or animal disease or harm. Researchers conducting experiments at DU are required to follow specific guidelines which are adapted from federal, state, and local agencies. The most effective means to minimize potential hazards associated with recombinant DNA and infectious agents is to combine professional experience with carefully performed laboratory procedures. Contact: Institutional Biosafety Committee – 303-871-2121 [email protected] Experimental protocols shall be submitted to the DU Institutional Biosafety Committee (IBC), via IRBNet, https://irbnet.org. Version 3.0 2020 4 1. BIOSAFETY GUIDELINES AND STANDARDS The CDC/NIH Biosafety in Microbiological and Biomedical Laboratories, 5th Edition, the OSHA Occupational Exposure to Bloodborne Pathogens Standard, and the NIH Guidelines involving Research with Recombinant DNA Molecules are most often referred to for the biomedical research that is conducted at DU. 1.1 CDC/NIH Biosafety in the Microbiological and Biomedical Laboratories The CDC/NIH “Biosafety in the Microbiological and Biomedical Laboratories” guidelines review the principles of biosafety including laboratory practices and techniques to be used in the biological laboratory conducting experiments with infectious agents and other potentially infectious materials. Agent summary listing and statements are provided, including description of practices to be followed for the more common infectious agents. The listing is categorized per the class of infectious agent, i.e. parasitic, fungal, bacterial/rickettsial, and viral. Safety equipment, facility design, biosafety levels and import and interstate shipment of biomedical materials are addressed. Practices and techniques to be followed at these biosafety levels are described. Animal biosafety levels are also described. The biosafety level classifications are described in Section 3. Guidelines for shipping specimens and infectious materials may be found in Section VI. 1.2 OSHA Occupational Exposure to Bloodborne Pathogens Standard Title 29, CFR Part 1910.1030 The OSHA Occupational Exposure to Bloodborne Pathogens Standard, necessitates that DU provide and adhere to a written policy concerning the handling of such materials. Per the OSHA standard, DU employees are evaluated for the potential for exposure to bloodborne pathogens. All employees at risk are provided training concerning the hazards, recognition of tasks which would lead to exposure, how to use personal protective equipment and engineering controls to minimize or eliminate this exposure, epidemiology regarding HIV, Hepatitis B, Hepatitis C, Hepatitis B vaccination information, as well as incident reporting and incident required follow-up. In accordance with the OSHA BBP Standard shall be available in any work area where there is a potential of exposure. The materials that can lead to exposure to bloodborne pathogens are identified below. This list identifies general categories of infectious materials most commonly associated with bloodborne pathogens. Version 3.0 2020 5 Human blood and other potentially infectious materials are defined as follows: • Human blood, human blood components, products made from human blood • Other Potentially Infectious Materials (OPIM), including: Human body fluids: semen, vaginal secretions, cerebrospinal fluid, synovial fluid, pleural fluid, pericardial fluid, peritoneal fluid, amniotic fluid, saliva in dental procedures, any body fluid that is visibly contaminated with blood, and all body fluids where it is difficult to differentiate if the fluid is contaminated. Any unfixed human tissue or organ, other than intact skin, from a human - living or dead. Materials containing HIV, HBV, and HCV: cell or tissue cultures, organ cultures and HIV, HBV, HCV - containing culture medium or other solutions; and blood, organs, and/or other tissues from experimental animals infected with HIV, HBV, and HCV. If you have questions relating to bloodborne pathogens and/or risk associated with occupational exposure, please the IBC at 1-2121. 1.3. NIH Guidelines for Research Involving Recombinant DNA Materials The NIH Guidelines should be followed whenever research protocols involve the use of recombinant DNA materials. This document provides information regarding the protocol approval process and methods to assist research investigators to perform experiments in a safe manner to protect the worker and minimize the risk of exposure to potentially biohazardous materials. The guidelines identify what types of experiments are exempt as well as specific directions and protocol approval requirements necessary for specific categories of experiments. The approval process begins with the review of the projects involving recombinant DNA molecules by the DU Institutional Biosafety Committee (IBC). Also included in the NIH recombinant DNA Guidelines are recommendations for physical and biological containment, shipment of materials, classification of infectious agents, and biosafety level descriptions for animals, plants, and infectious agents. This summary of the guidelines should be used only as a reference and should be used in addition to the full guidelines as published by the NIH/CDC. The guidelines should be referred to for specific definitions and clarifications. If further questions require resolution, these can be referred to the IBC. A current listing of infectious agents and their appropriate biosafety level, as determined by the NIH and CDC are included in Section 2. Version 3.0 2020 6 Experimental protocols shall be submitted to the DU Institutional Biosafety Committee (IBC), via IRBNet, https://irbnet.org. (Call 303-871-2121 for assistance). Summary of the NIH Guidelines for Research Involving Recombinant DNA Molecules Exempt Experiments Experiments are exempt from NIH recombinant DNA Guidelines if: a) they do not include the use of organisms or viruses (ex. PCR, Northern or Southern blotting), b) they involve DNA segments consisting entirely of single non-chromosomal source or manipulations of a viral DNA source with the intentional introduction of a deletion or mutation into a viral genome involving no foreign DNA, c) they consist entirely of DNA from a prokaryotic host including its indigenous plasmids or viruses (propagated in that host or transferred by well-known physiological means) (ex. E. Coli), d) they consist entirely of DNA from a eukaryotic host including its chloroplasts, mitochondria, or plasmids (excluding viruses) propagated in that host (ex. transformation of human cells with human DNA), e) they consist entirely of DNA from different species that exchange DNA by well-known physiological processes and no human pathogen is involved. (ex., Salmonella and E. coli), and f) the rDNA molecules used in the protocols contain <1/2 of any eukaryotic viral genome, are propagated in E. coli K12, Saccharomyces, Bacillus subtillis or B. lichenformis host-vector system, or are recombinant DNA molecules extrachromosomal elements of Gram positive organisms as listed in Appendix C- V of the NIH Guidelines, and are not included in a higher level category requiring additional
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