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risk management

Guidance for IBC Registrations

Required Information Prior to Writing The Application

Personnel (section 1e): Net IDs of all personnel working on the project including the PI and lab manager

Locations  (section 2a):

  • Room numbers and their Biosafety level
  • Date of the last inspection by the Biosafety Officer (BSO) for each room
  • Serial numbers of all Biosafety cabinets (BSC)/tissue culture hoods

If working with animals  (section 2b): The AR# provided by IACUC

Grant Information (section 3b):

  • Grant titles and funding agencies
  • The abstract page of any grants 

SECTION 3a

Two general types of responses can be submitted for project descriptions:

1. The abstract page of a grant

2. A general desciption of experimental design (overall goal and procedures) without focusing on hypothesis or specific aims. The following is an example of such a response:

  • Overall goal: My lab studies the effect of gene "X" on cellular function "y".
  • Procedures: Recombinant proteins will be produced in E. coli and analyzed by biochemical methods. We will utilize recombinant plasmids and viral vectors as a means of over-expressing gene "x" in the cell population being studied. Cells will be treated in culture with biological toxin "z" to assess changes in function "y". Cells will be transfered to mice via tail vein injection.

SECTION 4

4b1. Click here for further guidance on registration categories for research involving recombinant DNA.

4e questions 6  and 7

The known function(s) of the insert gene plays a vital role in the risk assessment process.  One or multiple functions of the gene could be potentially hazardous if expressed in humans.  This could occur, for example, as the result of a laboratory acquired infection with viral vectors.  Searching a database such as Entrez Gene would help determine the functions of a given gene.

The following image illustrates how to find Entrez Gene from the Pubmed search engine. After clicking on the gene of interest, scroll down to "GeneRIFS: Gene References into Function" to search for potential gene hazards.

4g Helper virus

A helper virus is a virus used to aid in the replication of a viral vector.  It is most often applied to adeno-associated virus (AAV) based vectors.  If you are using Retro/Lenti virus or Adenovirus based vectors, answer “No”.

4i.   Recombinant Propagation

Cells or organisms used to create or increase quantities of recombinant DNA (plasmids, viral vectors etc.) need to be disclosed in this section.  Typical experimental procedures involve expansion of plasmids in E. coli and use of mammalian packaging cell lines for creation of viral vector particles.

4i4.   Avoiding Aerosol Production During Purification

Purification of recombinants generally involves centrifugation which can aerosolize organisms.  There is minimal risk if plasmids are purified from small volumes of bacterial lysates by using sealed tubes and/or performing centrifugation with sealed rotors and closed lids in small bench top micro centrifuges.  Likewise, harvesting culture supernatants from viral packaging lines poses minimal risk when performed in a certified biosafety cabinent (BSC).  Centrifugation of concentrated suspensions of viral vectors is of greater concern.  Such procedures should be performed with sealed tubes and sealed rotors or safety cups. Conjunctivitis and respiratory infection may result from exposure to aerolized Adenovirus.  Personnel who are pregnant or immunocompromised are at increased risk for infection. The use of eye and respiratory protection should be considered.

4j.  Community Considerations

This section is critical for the IBC to be able to perform its responsibilities as found in Section IV-B-2-b of the NIH Guidelines for Research Involving Recombinant DNA (April 2002) and is highly scrutinized by the IBC during the review of applications. The IBC requires the PI to perform a risk assessment of possible hazards and negative consequences to lab personnel (section 4j1 and 4j2), individuals in the community (section 4j 3a), and the environment (section 4j 3b) from an exposure (laboratory aquired) or loss of containment (environmental release) of the recombinant, microbe, or infected cells or animals. 

Risk assesments for viral vectors: When performing a risk assessment, the PI should keep in mind that laboratory acquired infections with viral vectors may result in the expression of insert genes (named in section 4e) by the infected personnel. Guidance in performing a risk assessment for work with viral vectors can be obtained from Powerpoint presentations at the BSO website. The BSO can also be contacted for additional assistance.

Risk assessments for low risk agents (E. coli K12 derivatives and mammalian expression plasmids): The PI should disclose that research involving such agents pose some low level of risk rather than stating they are completely devoid of risk. The effect of insert genes and antibiotic resistance selection markers should be considered when assessing whether cloning experiments involving E. coli bacteria will result in altered virulence, increased ability to survive in the environment or withstand standard treatment with clinically relevant antibiotics.  While mammalian expression plasmids pose minimal risk, a worst case scenario exposure such as intravenous administration of plasmids may result in transient expression of insert genes by exposed personnel.


SECTION 5

5a.  Risk Groups (RG)

The National Institutes of Health (NIH) uses a classification scheme using a scale of 1-4 to assess the risk associated with microbes.  These assignments are based on the effect that an organism is likely to have on a healthy adult human.  It should be kept in mind that children, the elderly, and immunocompromised individuals are more susceptible to infection and are likely to be at greater risk.

Risk Group 1 (RG1)

Agents that are not associated with disease in healthy adult humans.

Risk Group 2 (RG2)

Agents that are associated with human disease which is rarely serious and for which preventive or therapeutic interventions are often available

Risk Group 3(RG3)

Agents that are associated withl serious or lethal human disease for which preventive or therapeutic interventions may be available (high individual risk, but low community risk)

Risk Group 4(RG4)

Agents that are likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available (high individual risk and high community risk)

Appendix B-Table 1.  Basis for the Classification of Biohazardous Agents by Risk Group (RG), NIH Guidelines for Research Involving Recombinant DNA Molecules, 2002

5a. Registering E. coli Used for Cloning and Protein Expression

  • Type:  bacteria
  • Risk group:  1
  • Agent:  Escherichia coli:  K12, DH5 alpha, and B strain can be selected from the menu 

5a2. Other strains, serotypes, and modifiers can be added in the textbox

A partially completed entry is available to assist with registering laboratory strains of E. coli and as a guide for completion of entries for other microorganisms.

5a.  Registering Viral Vectors

  • Type:  viruses
  • Risk group:  2
  • Agent: Adenoviral vector, Lentiviral vector, Retroviral vector

5a2. Strains, serotypes, and species of origin:  replication deficient Retrovirus (e.g. MMLV), Lentivirus (e.g. HIV, SIV, FIV), Adenovirus (e.g. human type 5)

For help assessing the virulence of recombinant viral vectors, please see the following powerpoint based viral vector training modules.

5b1.  Select Agents

Select agents are microbes and toxins identified by as having the potential to pose a severe threat to public health and safety.  The Centers for Disease Control and Prevention (CDC) regulates the possession, use, and transfer of these agents.  The CDC Select Agent Program oversees these activities and registers all laboratories and other entities in the United States that possess, use, or transfer a select agent or toxin in an amount above the maximum allowed.

A complete list of all select agents and toxins is found here.

5c1.  Experimental Procedures

To adequately answer this question, the PI should disclose the following:

  • Whether the agent will be maintained only in culture.
  • The maximum anticipated culture volume and concentration.  For bacteria, it may be noted how long the culture will be incubated e.g. overnight.
  • Other procedures to be performed with the organism.
  • Method of inoculating/infecting other cells, tissues, or organisms (if applicable)
  • Method of inactivation of lysis of the agent.  Be certain to note when this is done as well as the agent or method used.
  • Method of inactivating the microbe in both liquid (e.g. culture supernatants) and solid (e.g. pipets, culture dishes) waste.  Concentration of a disinfectant such as bleach and the final concentration of bleach should be included. 

NOTE:  Ideally, the agent should be treated with a suitable disinfectant in the laboratory and then autoclaved prior to removal from the laboratory.


Need more help?

For help with safety procedures, selecting personal protective equipment (PPE), choosing appropriate containment, writing safety protocols or to schedule a laboratory inspection, please contact the Biosafety Officer.

Daniel Eisenman, Ph.D.
Biosafety Officer
Phone: (843) 792-4304
Fax: (843) 792-0284
Pager: (843) 792-0590/12883
 
 
 

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