Lentivirus Safety

Standard Operating Procedures (SOP) for Working Safely with Lentiviral Vectors

The following personal protective equipment MUST be worn when working with Lentiviral vectors:
• Gloves (consider double-gloving depending on the procedures being performed)
• Lab Coat
• Goggles
• Face shield

Special Handling Procedures
1. Cells exposed to lentiviral vectors may not be removed from the laboratory for experimental purposes unless inactivated by approved procedures.
2. If you need to aerate cultures, it must be done slowly and in a manner that minimizes the potential for aerosol creation. This action must be carried out in a class II biological safety cabinet.

3. When pouring and pipetting samples, it must be done gently and slowly and must be carried out in a class II biological safety cabinet

4. Extra precautions must be taken when using sharps. Appropriate substitutes for sharp items must be used whenever they are available. No sharps (including needles and Pasteur pipettes) may be used for working with Lentivirus-infected cell cultures nor when harvesting virus pellets. Use plastic aspiration pipettes instead of glass Pasteur pipettes.

5. For Aspiration- Use a plastic vacuum flask with a second vacuum flask connected to it as a backup, with non-collapsible tubing capable of withstanding disinfection. To the second vacuum flask attach a hydrophobic and a HEPA filter (or combination filter) to ensure that nothing is sucked into the house vacuum system. These 3 items must be attached in series from the vacuum source in the hood or a vacuum pump

Decontamination/Clean-Up Procedures
All materials that have come into contact with Lentiviral vectors should be disinfected using a 1:10 bleach solution before disposal. Additionally, all work surfaces must be disinfected with a 1:10 solution of bleach once work is completed and at the end of the work day. (Note: A 15 minute contact time is required for decontamination)

Waste Disposal Procedures
1. Non-Sharp Waste- All cultures, stocks, and cell culture materials must be disinfected and autoclaved prior to being disposed of into a double red bag-lined biohazard box.

2. Sharps Waste-All needles, syringes, razors, scalpels, Pasteur pipettes and pipette tips must be disposed of in an approved, puncture resistant sharps container. Sharps containers must not filled more that 2/3 of their capacity.

Injury/Exposure Incident Procedures
1. Eye or Mucous Membrane Exposure from Splash or Aerosols- rinse a minimum of 15 minutes using eye wash and report the incident to your supervisor immediately.

2. Skin Contamination-Wash affected areas with soap and water for 15 minutes and report the incident to your supervisor immediately.

3. Needlestick and/or Sharps Exposure- Wash affected areas with soap and water for 15 minutes. Immediately notify your supervisor. He/she will complete the Post-Exposure Incident Report and submit it to the Department of Environmental Health and Safety within 24 hours of the exposure. Contact biosafety officer to arrange for appropriate medical attention.

Spill Response Procedures
The following steps must be taken when cleaning up a spill:
1. Stop, notify others and isolate the area!
2. Put on appropriate PPE (lab coat, gloves, eye and face protection).
3. Remove glass/lumps with forceps or scoop if applicable and place into a rigid, puncture resistant container.
4. Small spills-Place paper towels soaked in bleach directly on the spill and let soak for 20 minutes.
5. Wipe up area and discard towels in biohazard waste container.
6. Continue wiping area with paper towels soaked in bleach until the spill area is completely cleaned.
7. Discard all materials in biohazard waste container.
8. Wash hands thoroughly.