Recombinant or Synthetic Nucleic Acid Incident Spill Response and Reporting

Policy Statement

Recombinant or Synthetic Nucleic Acid Incident Spill Response and Reporting

Institutional Biosafety Committee Standard Operating Procedure #: 300.2

I. Purpose

In order to comply with federal reporting requirements and to ensure timely and appropriate follow-up, Principal Investigators shall immediately report exposures and releases involving recombinant or synthetic nucleic acid (r/sNA) molecules as well as violations of the NIH Guidelines for Research Involving Recombinant or Synthetic Nucleic Acid Molecules (NIH Guidelines, 2013) to the Dartmouth College Biological Safety Officer (BSO).

This policy defines immediate reporting procedures for incidents involving r/sNA molecules. Detailed procedures on response to other biohazard spills and exposures can be found in the Emergency Response and Biohazard Exposure Control Plan found on the Biosafety website: http://www.dartmouth.edu/~ehs/biological/policies_sops.html.

II. Definitions

According to the NIH Guidelines (Section I-B), recombinant or synthetic nucleic acids are defined as:

• i. molecules that a) are constructed by joining nucleic acid molecules and b) that can replicate in a living cell, i.e., recombinant nucleic acids;
• ii. nucleic acid molecules that are chemically or by other means synthesized or amplified, including those that are chemically or otherwise modified but can base pair with naturally occurring nucleic acid molecules, i.e., synthetic nucleic acids, or
• iii. molecules that result from the replication of those described in (i) or (ii) above.

III. Responsibilities

All personnel working with, and responding to incidents involving, recombinant or synthetic nucleic acids are responsible for adherence to this policy.

IV. Personnel Exposure Procedures

a. Exposures:

• Needlesticks or other percutaneous injuries from a contaminated sharp item
• Splashes to mucous membranes (eyes, nose, mouth)
• Bites/scratches from animals that have been exposed to any recombinant or synthetic nucleic acid material, whether or not the exposure leads to illness

b. Immediate Response:

• SKIN exposure: Immediately remove contaminated personal protective equipment or clothing and wash the contaminated area with an iodine solution or antibacterial soap and copious water for 15 minutes.
• EYE exposure: Flush the eye with water for at least 15 minutes at an eyewash station.

c. Notify PI or Supervisor.

If PI/supervisor is not available, immediately proceed to next step.

d. Medical Treatment

• During work hours, report to Occupational Medicine at DHMC from 7:30 a.m. to 4:30 p.m. (DHMC Faulkner Building, Level 4, near parking garage). Phone: (603) 653-3850.
• After-hours and weekends: report to DHMC Emergency Room.
• If transport assistance is needed or injury requires emergency treatment, contact Safety and Security at (603) 646-3333 or (603) 646-4000 from cell phone
• If emergency, call 911.

e. Reporting

• i. Notify the Biological Safety Officer (BSO) immediately.
• ii. The BSO will investigate the incident and notify the IBC Chair and EHS Director.
• iii. The PI will complete an internal Incident Report form and submit it to the BSO and Risk Management within 24 hr.
• iv. If the IBC Chair and BSO determine that the incident involves (nonexempt) r/sNA molecules, the BSO will submit an NIH incident report to the NIH Office of Biotechnology Activities within 30 days. Incidents occurring in BSL2 laboratories resulting in an overt exposure will be immediately reported to NIH OBA.
• v. Office of Risk Management and the Vice Provost for Research will also receive a copy of the incident report.

V. Environmental Release Procedures

a. Examples:

• Significant spill or release of any r/sNA molecules outside of containment equipment
• Theft, loss, or release of any r/sNA molecules into the environment, including escape or improper disposal of a transgenic animal or plant

b. Immediate Response

See attached Appendix A for spill response procedures.

c. Reporting

• Notify the BSO immediately via email or phone if not already contacted for assistance with spill clean-up.
• If the IBC Chair and BSO determine that the incident involves (non-exempt)r/sNA molecules, the BSO will submit an NIH incident report to the NIH Office of Biotechnology Activities within 30 days. Incidents occurring in BSL2 laboratories resulting in an overt release will be immediately reported to NIH OBA.

VI. Use of r/sNA Molecules Without Approval from the IBC

a. Research using recombinant or synthetic nucleic acid molecules as defined above and as described in Sections III-A, -B, -C and -D of the NIH Guidelines must be approved by the IBC prior to initiation.

b. Reporting

• Failure to obtain IBC approval prior to initiation of r/sNA research must be reported to the BSO immediately.
• BSO will prepare a detailed report for review by the IBC Chair and Vice Provost for Research. The report will be sent to the NIH OBA within 30 days.

Biosafety Officer:
Brenda Petrella, PhD
petrella@dartmouth.edu
(603) 646-9790
pager: 443-4305

Appendix A

I. Preparing for Spills

Most spills involving BSL2 organisms and viruses and r/sNA molecules can be handled by researchers. Supplies to clean a spill appropriately must be available in any lab that works with or stores biohazardous materials.

II. Recommended Supplies

a. Disinfectant

• Use appropriate disinfectant against the agents of concern, such as a fresh 1:10 bleach solution. Alcohol is not recommended for larger spills due to flammability concerns.

b. Personal Protective Equipment (PPE) (minimally)

• Lab coat
• Gloves
• Face shield or other mucous membrane protection

c. Other

• Sharps container for broken glass
• Absorbent pads
• Tongs
• Autoclave bags

III. Small Spills

Wipe up spill with a disinfectant-soaked paper towel and clean the surface with a suitable disinfectant. Allow disinfectant 20 minutes of contact time before removal.

IV. Large Spills

a. Spills outside of a containment device, i.e., the spill is not inside of a Biological Safety Cabinet (BSC), centrifuge, or other lab equipment

1. Close off spill area to traffic, and notify coworkers.
2. If the spill involved an aerosol, instruct all occupants to leave the room for 30 minutes to allow aerosols to settle.
3. Place a sign on the door warning staff not to enter the room due to a spill.
4. Remove contaminated lab coat or clothing and wash exposed skin.
5. Put on clean gloves and lab coat.
6. Prepare enough volume of a 1:10 dilution of chlorine bleach or other approved disinfectant to saturate the contaminated area.
7. Contain the spill with paper towels or other absorbent pads.
8. Flood the spill area with disinfectant. Leave on for 20 minutes.
9. Push the absorbent material at the edge of the spill into the spill's center. Add more paper towels as needed.
10. If glass is present, use tongs or forceps and a dustpan to remove pieces and place into a biohazard sharps container.
11. Discard the paper towels into an orange autoclave waste bag.
12. Using clean paper towels and a disinfectant, wipe all surfaces that may have come in contact with the spilled material.
13. Discard any disposable PPE into an orange autoclave waste bag.
14. Wash hands thoroughly.
15. Notify Principal Investigator or Supervisor and the BSO.

b. Spills inside of a Biological Safety Cabinet (BSC)

1. Leave BSC on.
2. Follow steps 4 through 15 above. Do not use 70% ethanol as it evaporates too quickly to allow adequate surface contact time.
3. If the cabinet has a catch basin beneath the work surface and the spill resulted in liquids flowing into this area, more extensive decontamination is required.
4. Ensure the drain valve under the cabinet is closed.
5. Pour disinfectant onto the work surface and through the front and rear grilles into the drain pan. Allow 20–30 minutes contact time.
6. Absorb spilled fluid-disinfectant from work surface with paper towels and discard in an orange autoclave bag.
7. Prepare to empty drain pan. Place fresh disinfectant solution into a collection vessel. Attach flexible tubing to the drain valve. The tube should be of sufficient length to allow the open end to be submerged in the collection vessel to minimize aerosol generation.
8. Open the drain valve and empty the drain pan into the collection vessel containing disinfectant. Flush the drain pan with water and remove the flexible tubing. Manage contaminated materials as if they are infectious.
9. Remove protective clothing used during cleanup and place in a biohazard bag for autoclaving.
10. Wash hands thoroughly.
11. Notify Principal Investigator or Supervisor and the BSO to determine if vapor/gas decontamination of the cabinet and filters is necessary.
12. Run BSC at least 10 minutes after cleanup, before resuming activity in the cabinet.

c. Spills Inside of a Centrifuge

Spills or breakage of containers inside of an operating centrifuge pose a serious potential for exposure due to the creation of aerosols. If a primary container has broken in a centrifuge without a closed rotor or bucket, immediately suspend use, notify lab staff and PI and request assistance from the Biosafety Officer. For suspected or confirmed spills/breakage in any centrifuge, wait at least 30 minutes after the centrifuge has stopped operating to initiate clean up.

1. Put on lab coat, gloves and a face shield prior to opening centrifuge. Open carefully to assess the damage.
2. If the spill is contained within a closed cup, bucket or rotor, spray the exterior with disinfectant and allow at least 20 minutes of contact time. Remove the carrier to the nearest biosafety cabinet (BSC). If a biosafety cabinet is not available, close the centrifuge; post a sign to indicate it cannot be used. Notify the PI and Biosafety Office for assistance.
3. If a BSC is available, gather supplies needed, such as a sharps container for broken glass and bins filled with disinfectant and place into the BSC. Use forceps to remove broken glass and place directly into sharps container. Carefully remove any unbroken tubes and place into a bin filled with disinfectant for 20 minutes. Wipe carrier/bucket with disinfectant.
4. After disinfection, the carrier, bucket or rotor should be washed with a mild soap and water.
5. Spray the interior of the centrifuge chamber with a disinfectant, let sit for 20 minutes and then wipe down.
6. Dispose of all clean-up materials (except sharps) in orange autoclave bags. Dispose of sharps in biohazard sharps containers.
7. Remove protective clothing and thoroughly wash hands.