FACT SHEET
Adenovirus

This fact sheet applies to any work with adenovirus, adenoviral vectors, and Adeno-Associated Virus (AAV). Work with these agents requires implementation of BSL-2 practices and procedures PLUS precautions detailed below (BSL-2 + Adeno). Prior to commencing any work with these agents, a protocol must be approved by the Institutional Biosafety Committee (IBC). Click here for information on the IBC registration/application process.

For a description of BSL-2 procedures, refer to CDC's Biosafety in Microbiological and Biomedical Laboratories (pages 20-27). PDF

BACKGROUND

• Adenovirus is a pathogen of respiratory and gastrointestinal mucous and eye membranes.

• Adenovirus (replication deficient and replication competent) can cause corneal and conjuctival damage. Eye protection (goggles) must be worn when working with this agent/vector.

• Adenovirus (unlike HIV or herpes) is quite stable. After having been extracted with ether and/or chloroform, it can still be infective.

• The replication-defective virus may be complemented in vivo thereby causing the vector to become replication competent.

MODE OF TRANSMISSION
Adenovirus may be transmitted by:

• Droplet

• Aerosol

• Injection

SYMPTOMS OF EXPOSURE
Any of these symptoms may occur following adenovirus exposure:

• Acute respiratory illness (cold like symptoms)

• Pneumonia

• Conjunctival infection (red eye)

• Corneal inflammation leading up to scarification

LABORATORY PRACTICES

  1. NO work with adenovirus is permitted on the open bench. A Biosafety Cabinet must be used for all manipulations including (but not limited to):
    • Pipetting
    • Harvesting infected cells for RNA
    • Loading and opening containers

    Exemptions will be considered on a case by case basis by the Biological Safety Officer and require approval by the Institutional Biosafety Committee.

  2. Within the Biological Safety Cabinet (BSC):
    • Plastic pipet tips and serological pipettes are to be placed in a cardboard box prior to disposal.
    • This box is placed into an autoclave bag with any other biohazardous waste in the BSC. The bag is to be closed and then sprayed with a disinfectant approved for efficacy against adenovirus (5% phenol or 10% household bleach).
    • The disinfected bag is then placed into a clean autoclave bag, and subsequently autoclaved for 1 hour at 121°C or 250°F (15 lbs per square inch of steam pressure).
  3. All vacuum lines must be fitted with a hydrophobic and HEPA filter. (ex. Fisher catalog Numbers: 09-744-75, SLFG 050 10, 09-730-211) Click here for an NIH example.
  4. Biohazard waste containers should be hard-sided with a foot operated lid, and easily decontaminated with liquid disinfectant. The autoclave bag should be folded over the rim so that only the inside of the bag is visible.
  5. All centrifugation must be done in closed containers using sealed rotors.
  6. Signs and labels must be placed to indicate each area where Adenovirus is used or stored (biological safety cabinets, incubators, refrigerators, laboratory entrance doors, etc.).
  7. Laboratory must have negative air pressure relative to the hallway (confirmed during lab audit).
  8. Laboratory gowns should be water resistant and have cotton knit cuffs with back closure and wrap around ties (ex. Fisher catalog numbers: 19 999 344A, and 17-988-496A).
  9. The most effective germicides (with a minimum 15 min. contact time) are:
    • Phenol (5%)
    • Sodium hypochlorite (household bleach diluted to 200 ppm or 10%).
  10. Adeno-associated viral vectors must be tested for presence of replication competent adenovirus after heat inactivation.
  11. Respiratory protection is necessary for adenoviral work. If experiment parameters make it impossible to work with adenovirus within a laminar flow biosafety cabinet, a respiratory protection program must be in place and N95 fit-tested respirators must be worn. Contact Occupational Health and Safety or the Biological Safety Officer for more information.
  12. Gloves must be worn at all times when working with adenovirus. Remove gloves using the inside-out technique. Dispose of gloves into biohazard waste container to be autoclaved. Wash hands immediately after removing gloves and before leaving work area. Never wear gloves outside of the laboratory, or touch doorknobs, telephones, personal belongings, etc. with gloved hands.
  13. All cultures, stocks, or materials used to manipulate or otherwise exposed to adenovirus must be autoclaved prior to disposal (except sharps). Autoclave conditions to be met: 1 hour at 121°C or 250°F (15 lbs per square inch of steam pressure). The outside of sharps buckets must be decontaminated before pick-up by Environmental Management for incineration.

ANIMAL USE

  1. Concurrent approvals are needed from UK Institutional Biosafety Committee and UK Institutional Animal Care and Use Committee prior to commencing animal work with adenovirus.
  2. Animals must be handled in a BSL-2 area designated and approved for adenoviral work.
  3. Infected animals may excrete (shed) adenovirus (especially the first 72 hours after infection). Precautions must be taken not to create aerosols when emptying animal waste material, washing cages, or cleaning the room.
  4. Special training must be given to all animal husbandry personnel on adenovirus. This training must address the hazards associated with the work, required practices and procedures and proper handling of bedding, cage washing, and all other husbandry materials associated with the experiment.
  5. All necropsy must be performed in a necropsy room using Animal BSL-2 +Adenovirus precautionary practices and procedures.
  6. Only lab personnel or animal husbandry workers trained to handle animals infected with adenovirus should be responsible for animal husbandry practices during the first 72 hours following infection of the animal.

EMPLOYEE EXPOSURE

Eye exposure from splash or aerosol:
Rinse a minimum of 15 minutes in eye wash or flush with water. Notify the Principal Investigator or Laboratory Supervisor, who will immediately contact Worker's Care at 1-800-440-6285 and direct the exposed employee to appropriate medical treatment at University Health Services, and to report the incident.

Needlestick, sharps exposure or non-intact skin exposure:
Contaminated skin should be scrubbed for approximately 20 minutes using a 10% povidone iodine solution (such as Betadine®) and copious amounts of water. Notify The Principal Investigator or Laboratory Manager, who will immediately contact Worker's Care at 1-800-440-6285 and direct the exposed employee to appropriate medical treatment at University Health Services, and to report the incident.

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