Overview of the NIH Guidelines and VT IBC

VT IBC; 3/2013

Institutional Biosafety Committee The IBC is involved in the establishment of biosafety policies that are used by Virginia Tech instructional and research personnel to safely conduct activities involving recombinant DNA, artificial gene transfer, infectious agents and biologically derived toxins.

The IBC ensures that all instructional and research laboratory activities are conducted in compliance with requirements and federally mandated guidelines, such as those outlined by the NIH, CDC, USDA, OSHA and Virginia Tech.

The IBC is a fundamental component of the NIH guidelines for research involving recombinant or synthetic nucleic acid molecules (NIH Guidelines).

NIH Guidelines – what are they? The NIH guidelines are a standard set of rules that are to be followed

by anyone working on research involving recombinant and/or synthetic nucleic acids, by specifying appropriate biosafety practices and procedures.

The guidelines outline the practices that must be followed when creating and handling recombinant and/or synthetic nucleic acids; as well as any organisms containing recombinant and/or synthetic nucleic acids.

a. Recombinant nucleic acids refer to DNA that is made outside of a living cell by ligating DNA sequence(s) to a replicative DNA molecule (i.e. plasmid DNA).

NIH Guidelines – Who is affected? The guidelines ensure that everyone shares the same understanding

regarding safety considerations, the types of experiments covered by the guidelines, and the roles and responsibilities of everyone associated with recombinant DNA and/or synthetic nucleic acids (rDNA) research.

The guidelines apply to any institution receiving NIH funding for rDNA research.

If rDNA research is not in compliance with the guidelines:a. NIH funding can be suspended or terminated for the non-

compliant project.b. NIH funding can be suspended or terminated for ALL NIH funded

projects that involve recombinant and/or synthetic nucleic acids at the institution.

c. All future projects involving recombinant and/or synthetic nucleic acids may require NIH approval prior to beginning any work.

Safety Considerations Risk Groups (RG) and biosafety containment levels (BL)

a. There are 4 levels of both risk groups and biosafety containment; RG-1-4 and BL-1-4. Level 1 is the lowest.

b. Risk Groups categorize biological agents (i.e. rDNA) by the potential risk associated with its use (NIH guidelines Appendix B).

c. Biosafety containment levels describe the specific equipment, practices and facilities that must be used for containment of biological agents (NIH guidelines Appendix G).

d. The RG designation may differ from the BL required.

Containment is both physical and biologicala. Physical includes techniques, equipment and facilities used to

contain the rDNA in the lab.b. Biological includes the survival and transmission ability of the

rDNA and organisms outside of the lab.

Safety Considerations – Risk Groups

Risk Group

Definition Examples

1Agents that are not associated with disease in healthy adult

humans B. subtilis

2Agents that are associated with human disease which is rarely

serious and for which preventive or therapeutic interventions are often available

Salmonella

3Agents that are associated with serious or lethal human disease

for which preventive or therapeutic interventions may be available (high individual risk but low community risk)

Prions, HIV types 1 and

2

4Agents 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)

Lassa virus, Ebola virus; NOT

permitted at Virginia Tech

NIH Guideline Experiments The NIH recognizes 6 rDNA review sections/categories:

1. Experiments requiring NIH director, RAC and IBC approval prior to initiation.

2. Experiments requiring IBC and NIH/OBA approval prior to initiation.

3. Experiments requiring IBC, IRB and RAC approval prior to participant enrollment.

4. Experiments requiring IBC approval prior to initiation.5. Experiments requiring IBC notice at initiation.6. Experiments exempt from the NIH guidelines.

Virginia Tech requires that all research involving recombinant and/or synthetic nucleic acids is registered with the VT IBC prior to any work being initiated, regardless of the NIH review category.

Section III-A

Experiments, considered to be Major Actions, that require RAC review, NIH Director approval and VT IBC approval.

Experiments involving the deliberate transfer of a drug resistance trait to microorganisms that are not known to acquire the trait naturally, if such acquisition could compromise the ability to control disease agents in humans, veterinary medicine, or agriculture, will be reviewed by the RAC.

Section III-B

Experiments that require NIH/OBA and VT IBC approval.

• III-B-1 regulates experiments involving the cloning of toxin molecules with an LD50 of less than 100 nanograms/kilogram of body weight.

•III-B-2 regulates experiments that have previously been approved as Major Actions (Section III-A) under the NIH Guidelines. Specific experiments already approved are included in Appendix D , “Major Actions Taken under the NIH Guidelines”.

Section III-C

Experiments that require VT IBC and VT IRB approvals s well as RAC review.

III-C-1 regulates experiments involving the deliberate transfer of recombinant or synthetic nucleic acid molecules, or DNA or RNA derived from recombinant or synthetic nucleic acid molecules, into one or more human research participants. No research participant shall be enrolled until the RAC review process has been completed

Section III-D

Experiments that require VT IBC approval.

• III-D-1 regulates experiments that involve the use of RG-2, RG-3, RG-4, or restricted agents as host-vector systems.

• III-D-2 regulates experiments in which DNA from RG-2, RG-3, RG-4, or restricted agents is cloned into non-pathogenic prokaryotic, or lower eukaryotic, host-vector systems.

• III-D-3 regulates experiments involving the use of infectious DNA or RNA viruses, or defective DNA and/or RNA viruses in the presence of helper virus, in tissue culture systems.

Section III-D, continued

• III-D-4 regulates experiments involving whole animals in which the animal's genome has been altered by stable introduction of recombinant or synthetic nucleic acid molecules, or nucleic acids derived therefrom, into the germ-line (transgenic animals), as well as experiments involving viable recombinant or synthetic nucleic acid molecule-modified microorganisms tested on whole animals, which must be conducted using a minimum containment level of BL-2.

• III-D-5 regulates experiments to genetically engineer whole plants by recombinant or synthetic nucleic acid molecule methods, to use such plants for other experimental purposes (e.g., response to stress), to propagate such plants, or to use plants together with microorganisms or insects containing recombinant or synthetic nucleic acid molecules; if the experiments involve agents requiring BL-2 containment.

Section III-D, continued

• III-D-6 regulates experiments involving the generation of more than 10 Liters of culture, at any one time.

• III-D-7 regulates experiments involving Influenza viruses.

Section III-E

Experiments requiring VT IBC approval.

III-E regulates experiments not included in Sections III-A, B, C, D, F, and/or their subsections.

• III-E-1 regulates experiments involving the formation of recombinant or synthetic nucleic acid molecules containing no more than 2/3 of the genome of any eukaryotic virus.

• III-E-2 regulates experiments involving nucleic acid molecule-modified whole plants, and/or experiments involving recombinant or synthetic nucleic acid molecule-modified organisms associated with whole plants, except those that fall under Section III-A, III-B, III-D or III-F. These experiments do not involve the use of exotic plants and/or noxious weeds.

Section III-E, continued

• III-E-3 regulates experiments involving the generation of rodents in which the animal's genome has been altered by stable introduction of recombinant or synthetic nucleic acid molecules, or nucleic acids derived therefrom, into the germ-line (transgenic rodents). Only experiments that require BL1 containment are covered under this section; experiments that require BL2, BL3, or BL4 containment are covered under III-D-4, “Experiments Involving Whole Animals”.

Section III-F

Experiments requiring VT IBC approval.

III-F regulates experiments involving recombinant or synthetic nucleic acid molecules that are exempt from the NIH Guidelines.

• III-F-1: Involving synthetic nucleic acids that: (1) can neither replicate nor generate nucleic acids that can replicate in any living cell (e.g., oligonucleotides or other synthetic nucleic acids that do not contain an origin of replication or contain elements known to interact with either DNA or RNA polymerase), and (2) are not designed to integrate into DNA, and (3) do not produce a toxin that is lethal for vertebrates at an LD50 of less than 100 nanograms per kilogram body weight.

• III-F-2: Involving rDNA that are not in organisms, cells, or viruses and that have not been modified or manipulated to render them capable of penetrating cellular membranes.

Section III-F, continued

• III-F-3: Involving rDNA that consist solely of the exact recombinant or synthetic nucleic acid sequence from a single source that exists contemporaneously in nature.

• III-F-4: Involving rDNA that consist entirely of nucleic acids from a prokaryotic host, including its indigenous plasmids or viruses when propagated only in that host (or a closely related strain of the same species), or when transferred to another host by well-established physiological means.

•III-F-5: Involving rDNA Those that consist entirely of nucleic acids from a eukaryotic host including its chloroplasts, mitochondria, or plasmids (but excluding viruses) when propagated only in that host (or a closely related strain of the same species).

Section III-F, continued

• III-F-6: Involving rDNA that consist entirely of DNA segments from different species that exchange DNA by known physiological processes, though one or more of the segments may be a synthetic equivalent.

• III-F-7: Involving genomic DNA molecules that have acquired a transposable element, provided the transposable element does not contain any recombinant and/or synthetic DNA.

• III-F-8: Involving rDNA experiments that do not present a significant risk to health or the environment (see Appendix C).

Responsibilities of the VT Principal Investigator (PI)

In regards to IBC protocol submissions, the PI shall • Make an initial determination of the risk groups and containment levels

required to safely conduct the research in the protocol.• Select the appropriate microbiological and lab techniques.• Submit the initial protocol, as well as subsequent changes, to the IBC

for review and approval or disapproval.• Maintain active communication with the IBC throughout the conduct of

the research.Prior to beginning the research (after IBC approval), the PI shall

• make all protocols and lab manuals available to all personnel, describing the potential biohazards of the work.

• Instruct and train all personnel in practices and techniques required to ensure safety; and in the procedures for dealing with accidents.

• Maintain written documentation of personnel training.• Inform all personnel of the reasons and provisions for any

precautionary medical practices that are advised or requested (e.g. vaccinations, serum collection).

Responsibilities of the VT PI (continued) During the conduct of the research, the PI shall

• Supervise the safety performance of all personnel, to ensure that the required safety practices and techniques are employed.

• Investigate and report any potential exposure, accident, or incident pertaining to the operation and implementation of containment practices and procedures

• The Biosafety Officer must be informed, immediately• The following people/departments must be notified, in

writing: Biosafety Officer, VT IBC, greenhouse/animal facility manager, NIH/OBA (the IBC and BSO will help the PI submit this report).

Correct work errors and conditions that may result in the release of recombinant and/or synthetic nucleic acid molecules.Ensure the integrity of physical and biological containment in the lab(s).Comply with reporting requirements for human gene transfer experiments.

VT IBC The purpose of the VT IBC is to ensure the health and safety of all personnel working with biohazardous agents.

VT research and teaching faculty are responsible for registering all work involving biohazardous agents. Information regarding the biohazardous agents overseen by the VT IBC can be found on our website (http://ibc.researchcompliance.vt.edu/), or by contacting the IBC administrator at IBC@vt.edu.

References

VT Institutional Biosafety Committee (IBC) website

NIH/OBA website

NIH Guidelines (March 2013)

Quiz

Please complete the required online quiz, using the following link https://auth.vt.edu/login?service=https%3A%2F%2Fsecure.research.vt.edu%2Forc_cert%2Findex.php%3Fmod%3Dquiz