Johns Hopkins Biosafety Manual 1995
JOHNS HOPKINS INSTITUTIONS.
BIOSAFETY MANUAL.
TABLE OF CONTENTS.
June 1995. A trip down memory lane.
Full Text available at researchgate.net.
FORWARD.
The Johns Hopkins Hospital and the Johns Hopkins University have high standards in teaching and research. The University and Hospital are committed to provide an academic and working environment as free as possible from all hazards. If we are to honor our commitment we must have the continued cooperation of faculty, administration, research and supervisory personnel for the establishment of safe procedures and for providing good training and adequate facilities for the protection of employees, students, patients and visitors.
The administration of the safety program has been placed with the Office of Safety and Environmental Health which offers a wide range of technical assistance, training aids and reference materials in biohazard control. Please use these resources to implement your safety program.
Edward J. Bernacki, M.D., M.P.H., Chair.
Joint Committee on Health, Safety and Environment.
PREFACE.
This Biosafety Manual was prepared to provide laboratory and clinical investigators current information on biohazard control. The manual will be updated as our knowledge of the risks involved in the handling of biological hazards increases and as methods for containment of these hazards improve.
The Biosafety Manual brings together information which will assist supervisors in carrying out their responsibilities in the management of biohazards. The Manual provides suggested procedures, techniques and equipment to protect personnel, animals and experiments from unintentional infection or contamination in addition to pertinent policy generated by Hopkins committees and standards resulting from federal, state and local legislation.
The manual represents a compendium of biohazards control information that can serve as a base for specific programs. All personnel using potentially biohazardous materials should become familiar with the Biosafety Manual and should conduct their operation in accordance with the level of risk appropriate for the procedure and the potentially biohazardous agent or materials they handle.
The success of the biohazardous control program depends on investigators who are motivated toward a safe working environment and who have knowledge of safe operational procedures. The Biosafety Office will endeavor to do its part by providing accurate information and technical assistance to assist in the establishment of adequate biohazards controls for the protection of both the investigators and their experiments.
Richard W. Gilpin, Ph.D., R.B.P.
Biosafety Officer.
Byron S. Tepper, Ph.D., C.S.P.
Director, Office of Safety & Environmental Health.
TABLE OF CONTENTS.
SECTION I — SAFETY RESPONSIBILITIES.
Administrative Responsibilities…………………………………..…………1
Administrative Officers………………………………………………………1
Department Chairman and/or Director……………………….………….1
Principal Investigator and/or Supervisor……………………..…………1
Individual Researchers and/or Technicians…………………………….2
Periodic Reviews and Updates of Registered Research…..………….3
Biological Agents and/or Materials……………………………………….3
Recombinant DNA………………………………………………….…………3
Registration of Research with
Pathogenic and/or Oncogenic Materials……………………..…………4
Background………………………………………………………………………4
Definition…………………………………………………………………………4
Responsibility………………………………………………………..…………4
Assessment of Hazard……………………………………………..…………5
Purpose of Registration………………………………………………………5
Procedure for Registration………………………………………..…………6
Standard Operating Procedure (SOP) Manual
for Biosafety Level 3 Laboratories…………………………..…………7
Vaccinia Virus Vaccination………………………………………..………….8
Registration of Research with HBV and/or HIV……………………….9
Requirements……………………………………………………………………9
Informed Consent for Use of Postexposure
Zidovudine (AZT)…………………………………………………………..…9
Registration of Research with Sheep or Goats……………..…………12
Background………………………………………………………….…………12
Recommended Control Measures for Q Fever………………………12
Registration of Research with Recombinant DNA……….…………14
Registration Form……………………………………………………………14
Review Procedure……………………………………………………………14
Review of Recombinant DNA Research With
Moderate Risk Biological Agents or Materials
at Biosafety Level 1 or 2…………………………………………………..15
Review of Recombinant DNA Research Involving
Biosafety Level 3 for High Risk Biological Agents
or Materials or Recombinant DNA Involving Human Subjects..15
Registration of Research with Non-Human Primates……………….17
Tuberculosis……………………………………………………………………17
Herpesvirus-B in Macaque Monkeys……………………….…………18
SECTION II — SAFETY COMPLIANCE.
Standard Practices, Training and Precautions…………………………21
Universal Precautions Guidelines for Research Laboratories..……23
Definitions…………………………………………………………..…………23
Precautions…………………………………………………………..…………23
HBV and/or HIV Agent Summary Statements…………….………….27
Q Fever Precautions………………………………………………..…………28
Background………………………………………………………….…………28
Rationale……………………………………………………………..…………28
Registration………………………………………………………….…………28
Medical Surveillance……………………………………………..…………29
Transport of Animals…………………………………………….………….29
Personnel Protection…………………………………………………………30
Waste Disposal…………………………………………………….………….31
Decontamination…………………………………………………..…………31
Biosafety Level 1 Guidelines………………………………………………32
Biosafety Level 2 Guidelines………………………………………………33
Biosafety Level 3 Guidelines………………………………………………34
Animal Biosafety Levels………………………………………….…………35
Biosafety References……………………………………………….…………36
Biosafety Level 3 Facility Specifications………………………………38
Performance……………………………………………………………………38
Location…………………………………………………………………………38
Floors………………………………………………………………….…………38
Walls…………………………………………………………………..…………38
Ceiling………………………………………………………………..…………38
Access Zone……………………………………………………………………38
Doors………………………………………………………………….…………38
Windows……………………………………………………………..…………39
Plumbing……………………………………………………………..…………39
Vacuum System………………………………………………………………39
Electrical……………………………………………………………..…………39
Light Fixtures……………………………………………………….…………39
Lab Furniture……………………………………………………….…………39
Hand Washing Sink…………………………………………………………40
Eye, Face and Body Spray Fixture…………………………..…………40
Autoclave…………………………………………………………….…………40
Ventilation…………………………………………………………..…………40
Biological Safety Cabinets……………………………………..…………41
Chemical Fume Hood…………………………………………..………….41
Air Pressure Differential……………………………………….………….41
Directional Airflow……………………………………………….…………42
Air Supply Vents…………………………………………………..…………42
Fire Protection…………………………………………………………………42
Summary of Biosafety Levels…………………………………..…………43
Classification of Etiologic Agents on the Basis of Hazard………..44
Classification of Ongogenic Viruses on the Basis of Hazard……..45
Criteria for Moderate Risk Oncogenic Viruses…………..…………45
Criteria for High Risk Oncogenic Viruses……………………………45
General Laboratory Practices and Procedures……………..…………46
Employee Action………………………………………………….…………46
Food, Drink, and Cosmetics………………………………………………46
Hand Washing…………………………………………………………………46
Beards and Long Hair………………………………………………………47
Books and Journals……………………………………………….…………47
Pipettes………………………………………………………………..…………47
Syringes…………………………………………………………………………48
Disposal of Research Laboratory Waste……………………..…………49
Definition of Sharps…………………………………………………………49
Disposal of Laboratory Waste……………………………………………49
Approved Laboratory Waste Containers…………………..………….49
Utilization of Six or Ten Quart Plastic Containers…………………50
Utilization of the Biohazard Box……………………………..…………50
Additional Containers………………………………………………………51
Safe Disposal of Sharps Containers………………………….…………51
Disposal of General Laboratory Glass…………………………………52
Summary of Waste Disposed………………………………….………….52
Centrifuge Containment…………………………………………..…………53
Centrifuge Precautions…………………………………………..…………53
Centrifuge Tubes…………………………………………………..…………53
High-Speed Centrifuges……………………………………………………54
Containment Centrifugation………………………………………………54
Blenders, Sonicators, Mills, Grinders & Cell Sorters…….………..57
Water Bath Disinfectants……………………………………………………58
Laboratory Vacuum Traps……………………………………….………….59
Refrigerators, Freezers and Dry Ice Chests………………….…………61
Transportation of Biohazardous Material………………………………62
Test Tube and/or Vacutainer Techniques……………………………….63
Membrane Filters………………………………………………………………64
Hazardous Operations (Two Person Rule)………………….………….65
Use of Dry Powders………………………………………………..…………66
Laboratory Construction and Renovation………………………………67
Animal Care and Handling……………………………………….…………68
Decontamination and Disinfection Procedures…………….…………69
Steam Autoclaving………………………………………………….…………71
Autoclave Decontamination Procedures……………………..…………72
Background………………………………………………………….…………72
Indicators…………………………………………………………….…………72
Containers………………………………………………………………………74
Autoclave Operation……………………………………………..………….75
Processing Times………………………………………………….…………76
Odor Control………………………………………………………..…………77
Training…………………………………………………………………………77
Record Keeping………………………………………………………………77
Autoclave Log Book Samples……………………………………………78
Dry Heat Sterilization……………………………………………..…………83
Ethylene Oxide Sterilization…………………………………….…………84
Formaldehyde Gas Decontamination of Equipment……..…………85
Ultraviolet Light Decontamination…………………………….…………86
UV Lamp Operation…………………………………………………………86
Training…………………………………………………………………………87
Chemical Disinfectants………………………………………………………88
Phenolic Compounds…………………………………………….…………88
Quaternary Ammonium Compounds………………………..…………88
Iodophors…………………………………………………………….…………89
Alcohols…………………………………………………………………………90
Aldehydes………………………………………………………………………90
Chlorine Compounds…………………………………………….…………91
Mercurials………………………………………………………………………91
Clean-up of Biohazardous Spills……………………………….…………93
Biohazard Spill Inside a Biological Safety Cabinet…….…………93
Biohazard Spill Outside a Biological Safety Cabinet…..…………93
Radioactive Biohazard Spill Outside a
Biological Safety Cabinet…………………………………..….………..95
Shipment of Biological Materials……………………………………….…97
SECTION III — RECOMBINANT DNA GUIDELINES.
Introduction………………………………………………………..……….…..101
Table of Contents………………………………………………..……………101
Section I. Scope of the NIH Guidelines………………..……………104
Section II. Containment……………………………………………………106
Section III. Experiments Covered by the NIH Guidelines.……….107
Section IV. Roles and Responsibilities…………………..……………114
Section V. Footnotes and References of Sections I Through IV…124
Appendix A. Exemptions Under Section III-E-5
Sublists of Natural Exchangers………………………………………126
Appendix B. Classification of Etiologic Agents and
Oncogenic Viruses on the Basis of Hazard………………………..…..127
Appendix C. Exemptions Under Section III-E-6……..………….. .133
Appendix D. Major Actions Taken Under the NIH Guidelines…. 136
Appendix E. Certified Host-Vector Systems…………..………….…136
Appendix F. Containment Conditions for Cloning of Genes
Coding for the Biosynthesis of Molecules Toxic for Vertebrates..138
Appendix G. Physical Containment……………………….……………139
Appendix H. Shipment………………………………………..……………151
Appendix I. Biological Containment…………………….……………152
Appendix J. Biotechnology Research Subcommittee…………….155
Appendix K. Physical Containment for Large Scale Uses
of Organisms Containing Recombinant DNA Molecules………..155
Appendix L. Release into the Environment of Certain Plants…….165
Appendix M. Points to Consider in the Design and Submission
of Protocols for the Transfer of Recombinant DNA Molecules
into the Genome of One or More Human Subjects…………………..165
Appendix P. Physical and Biological Containment for
Recombinant DNA Research Involving Plants………………………..176
Appendix Q. Physical and Biological Containment for
Recombinant DNA Research Involving Animals…………………….187
SECTION IV — BIOHAZARD CABINETRY
Biohazard Containment Equipment………………………….…………202
Types of Containment Equipment……………………………………..202
Use of Containment Equipment…………………………….………….202
Biohazard Cabinet Containment Equipment…………….…………203
Biological Safety Cabinet Categories……………………….…………205
Use of Biological Safety Cabinets………………………………………208
Operation of Biological Safety Cabinets…………………..…………210
General Suggestions…………………………………………….…………210
Starting Up Your BSC…………………………………………………….210
Cleaning Your BSC…………………………………………….………….213
Flammable or Explosive Materials within BSC’s……..………….214
BSC Assignment for Specific Biosafety Levels…………………….215
Biosafety Cabinet Training Materials……………………….…………216
Characteristics of BSC’S and other Airflow Equipment…………218
Class 1 Biological Safety Cabinets……………………………………219
Class II, Type A Biological Safety Cabinets…………….…………221
Class II, Type B1 Biological Safety Cabinets…………..…………223
Class II, Type B2 Biological Safety Cabinet………………………226
Class II, Type B3 Biological Safety Cabinets…………..…………228
Class III Biological Safety Cabinets……………………….…………230
Other Cabinets………………………………………………………………232
Horizontal Laminar Flow Clean Air Benches (CAB’s)…………232
Chemical Fume Hoods……………………………………………………234
Biological Safety Cabinet Use Guidelines……………………………236
Purchasing Procedures for Biological Safety Cabinets..…………237
Vendors of Biological Safety Cabinets……………………..…………242
Biological Safety Cabinet Installation Guidelines…………………244
Set-up Location……………………………………………………………..244
Gas and Vacuum Utility Connections…………………….…………246
Electrical Service Connections…………………………………………246
New Installations……………………………………………………………247
Working Height………………………………………………….…………247
Thimble Connection Exhaust Air Venting………………..………….248
Hard Connection Exhaust Air Venting……………………..…………255
Frequency of Cabinet Certification………………………….…………257
Annual Cabinet Labor and Service Contract……………..…………258
Scheduling Cabinet Service and Certification………………………259
Moving Your Cabinet……………………………………………………….261
Certification Tests……………………………………………………………262
SECTION V — HAZARD WARNING SIGNAGE.
Hazard Signage…………………………………………………….…………265
Hazard Warning Labels…………………………………………………….269
Caution Signage
Biohazard, Infectious Agents, BL2……………………………………278
Biohazard, Potentially Infectious Material, BL2……….…………280
Biohazard, Infectious Agents, BL3……………………………………282
Biohazard, Infected Animals……………………………………………284
Radioactive Materials…………………………………………..…………286
Radiation Area………………………………………………………………288
High Radiation Area………………………………………………………290
Hazardous Chemical, Cancer Suspect Agent………………………292
Cancer Hazard…………………………………………………….…………294
Eye Protection Required……………………………………….…………296
Danger, Do Not Enter, Contaminated Area……………..………….298
Multiple Hazards……………………………………………………………300
ALPHABETICAL INDEX………………………………………….………303