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Live Virus Worker Web Training





training disclaimer


The use and handling of biological materials in the research setting always requires consideration for the potential of exposure to infectious agents, and how any exposure potential can be reduced or eliminated.  When live viruses are used in the laboratory, there is a need for even further care and consideration of exposure hazards since many viruses are related to human or animal disease.  For those working with live viruses at Princeton University, a special program has been established to ensure that appropriate training, instruction, and medical consultation are provided.  This web-based training is designed to provide you with the initial instruction and guidance on biosafety issues and help you fulfill the additional requirements for placement into the Live Virus Worker Program.  This presentation will conclude with a test to gauge your understanding of the information provided.  Upon successful completion of the test, you will be directed to information and forms to complete the Live Virus Worker application and medical review.

SPECIAL NOTE:  Those working with non-human retroviruses or insect viruses (i.e. baculovirus) are not required to be in the Live Virus Worker Program.


Biosafety Requirements and Guidelines

In the late ‘70s when work with recombinant DNA (rDNA) was first proposed, there was a certain amount of anxiety and hysteria about the biological hazards that would be unleashed by this type of research.   It became clear that there would have to be guidelines established that would ensure the safe use of these materials.  The result was the establishment by the National Institutes of Health (NIH) of the Guidelines for Research Involving Recombinant DNA Molecules (NIH Guidelines).  These national guidelines call for institutions doing rDNA research to establish an Institutional Biosafety Committee (IBC) to oversee research and provide the parameters under which rDNA work is to be conducted.  Those institutions receiving grant funds from NIH, as Princeton University does, are required to meet the provisions of the NIH Guidelines.  The NIH Guidelines have been revised and modified over the years to address work with plants, animals, human genome transfer, and large scale production involving rDNA.

BMBLSeveral years after the rDNA guidelines were established, an additional joint guidance document was provided from the Centers for Disease Control and Prevention (CDC) and NIH titled, Biosafety in Microbiological and Biomedical Laboratories(BMBL).  The BMBL expands on the biosafety guidelines established for rDNA work to include work with all microbiological agents, i.e., bacterial, fungal, parasitic, viral, rickettsial, and prions.  It has also been revised several times over the years to incorporate Biosafety Levels for work with biologically infected animals and several appendixes dealing with issues of handling and use of various biological materials.

Princeton University has adopted both of these guidelines as the basis of its policy for proper use and handling of potentially biohazardous materials.  Those working with biological materials, including live viruses, are expected to follow the biosafety measures which are part of these guidelines.

Biosafety Levels

The biosafety parameters in both the NIH and the CDC/NIH Guidelines are based on Biosafety Levels (BSLs).  The BSLs are the practices, procedures, equipment, and facilities needed to be in place depending on the hazard associated with the biological material or agent in use.  The BSLs move from the least restrictive conditions of BSL1 (basic good microbiological practice) to those of BSL4 which are needed for work with highly exotic agents.

By University policy, biological research at Princeton has been limited to work with BSL1 and BSL2 agents.

Biological agents considered to be at the minimal hazard level of BSL1 are not known to generally cause disease in healthy adults.  BSL2 agents are associated with human disease through exposures involving skin injury, ingestion, or mucous membrane contact.

The following table provides a brief synopsis of the requirements of each of the biosafety levels (BSL1 through BSL4).

BSL Agents 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 hand sink required
2 Associated with human disease, hazard = percutaneous injury, ingestion, mucous membrane exposure BSL-1 practice plus:
  • Limited access
  • Biohazard warning signs
  • "Sharps" precautions
  • Biosafety manual defining any needed waste decontamination or medical surveillance policies
Primary barriers = Class I or Biological Safety cabinet or other physical containment devices used for all manipulations of agents that cause splashes or aerosols of infectious materials; Personal Protective Equipment: laboratory coats; gloves; face protection as needed. BSL-1 plus:
Autoclave available
3 Indigenous or exotic agents with potential for aerosol transmission; disease may have serious or lethal consequences BSL-2 practice plus
  • Controlled access
  • Decontamination of all waste
  • Decontamination of lab clothing before laundering
  • Baseline serum
Primary barries = Class I or II BSC's or other physical containment devices used for all open manipulations of agents; PPEs: protective lab clothing; gloves; respiratory protection as needed BSL-2 plus;
  • Physical separation from access corridors
  • Self-closing, double-door access
  • Exhausted air not recirculated
  • Negative airflow into laboratory
4 Dangerous/exotic agents which pose high risk of life-threatening disease, aerosol-transmitted lab infections; or related agents with unknown risk of transmission BSL-3 practice plus:
  • Clothing change before entering
  • Shower on exit
  • All material decontaminated on exit from facility
Primary barriers = All procedures conducted in Class III BSC's or Class I or II BSC's in combination with full-body, air-supplied, positive pressure personnel suit BSL-3 plus:
  • Separate building or isolated zone
  • Dedicated supply and exhaust, vacuum, and decon systems
  • Other requirements outlined in the text of the BMBL

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The Biohazard Warning Symbol 

biohazard The biohazard symbol as shown here is a visual representation and reminder that viruses or other microbiological materials are present which may pose an exposure hazard resulting in illness.  You will find this symbol on biohazard signs posted where BSL2 level agents are being used, on containers and equipment in which biological materials are maintained, and on waste containers, such as sharps disposal boxes and medical waste boxes provided in the laboratory.

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University Biosafety Manual

The Biosafety Manual is available as a resource and reference document for information, guidelines, policies, and procedures to enable and encourage those handling and working with potentially biohazardous materials to work safely and reduce the potential for exposure. (This link may be viewed at the conclusion of the Live Virus Worker Training)

In the Manual, you can find information covered in this training about work with live viruses as well as other aspects of the Biological Safety Program, including:

  • The University’s Institutional Biosafety Committee (IBC)
  • Contacts and Responsibilities for Biosafety
  • Research Project Registration and Approval
  • Work with Blood and Human Tissue, and Infected Lab Animals
  • Safe Laboratory Practices and Procedures
  • Biosafety Cabinets
  • Decontamination
  • Biological Spills
  • Biological Waste
  • Packaging, Shipping, Import, and Export of Biological Materials
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Institutional Biosafety Committee

Princeton University’s  Institutional Biosafety Committee (IBC) was established in the late ’70s to comply with the NIH Guidelines and provide an oversight body operating under the University Research Board to:

  • oversee the biological safety program
  • review research proposals involving rDNA and other potentially biohazardous agents and approve those that comply with NIH and CDC guidelines and University policy
  • adopt policies supporting the safe use of biological materials and the elimination or reduction of exposure to potentially bio-hazardous materials or agents
  • address bio-safety issues related to experimentally-infected laboratory animals

peopleThe IBC is composed of faculty and administrative staff, and also includes individuals from Princeton Township and Borough who represent the interests of their communities.
The Committee is chaired by a faculty member and the Secretary is from the Office of Research and Project Administration (ORPA).  The University Biosafety Officer from Environmental Health and Safety (EHS), the Campus Veterinarian, a physician from the University Health Services, and faculty representing appropriate areas of expertise are also part of the Committee.  A list of the current Committee membership is found in the University Register.

The Committee has an initial meeting each Fall to consider new and review existing research proposals for the academic year.  Subsequent meetings are scheduled as needed to address biosafety issues and review any new proposals.

If you are involved in new research, ensure that it has been reviewed and approved by the IBC.

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