INSTITUTIONAL BIOSAFETY COMMITTEE
SAN DIEGO STATE UNIVERSITY
GUIDELINES
FOR RESEARCH WITH ADENOVIRUS
I.Introduction
This document provides essential information for Research Investigators regarding work with
human Adenovirus, Adenovirus vector systems and Adeno-Associated Virus (AAV) and vector
systems in the lab and in animal models. The IBC will determine the final Biosafety Level (BSL)
and/or Animal Biosafety Level (ABSL) for work with Adenovirus, Adenovirus vector system, and
AAV based on risk assessment and in accordance with the CDC/NIH Biosafety in Microbiological
and Biomedical Laboratories (BMBL), NIH Guidelines for Research Involving Recombinant DNA
Molecules (NIH Guidelines), and this SDSU IBC Guidelines for Research with Adenovirus. Specific
requirements within these guidelines will be explicitly indicated in the BUA Approval Letter. Use the
requirements specified in the BUA Approval Letter and indicated in the BUA Application as a
training tool to inform all research personnel of the procedures for working with Adenovirus,
Adenovirus vector system, and/or AAV and AAV vector systems in the research laboratory or
animal facility.
II.Adenovirus
Adenovirus is a human pathogen with 8 serotypes: 1, 2, 3, 4, 5, 7, 40 and 41 and classified in the
NIH Guidelines as a Risk Group 2. Adenoviruses most commonly causes respiratory illness;
however, depending on the infecting serotype, they may also cause various other illnesses, such as
gastroenteritis, conjunctivitis, cystitis (bladder infection), and rash illness. While occupational
exposure to Adenovirus is generally through aerosols, fomite contact, fecal-oral route and
percutaneous exposure are also exposure routes. Adenovirus is susceptible to some disinfectants
(Section IX.), but if work surfaces are not correctly disinfected the virus can survive for prolonged
periods outside of host body, in some cases 8 weeks on environmental surfaces at room
temperature. There is a vaccine for Adenovirus serotype 4 and Adenovirus serotype 7. The SDSU
Institutional Biosafety Committee (IBC) will determine the BSL of work involving Adenovirus based
on risk assessment. IBC has determined that work with any Adenovirus serotypes at a minimum
require BSL 2 facility and containment PLUS precautions specific to work with adenovirus (BSL-2 +
Adeno) as described in Section V of this document. Requirements for BSL 2 facility and work
practices are indicated in the BMBL Section IV – Laboratory Biosafety Level Criteria.
III.Adenovirus Vectors
The SDSU Institutional Biosafety Committee (IBC) will determine the BSL of work involving the use
of Adenoviral vectors based on NIH Guidelines and risk assessment. The risk assessment will
evaluate the potential for generation of replication/competent adenovirus (RCL), aerosolizing
procedures and the potential for oncogenesis from the transgene among other risk factors.
Work with adenovirus in the vector form will be at a minimum BSL 2 facility and work practices
PLUS precautions specific to work with Adenovirus (BSL-2 + Adeno) as described in Section V of
this document. Requirements for BSL 2 facilities and work practices are indicated in BMBL Section
IV – Laboratory Biosafety Level Criteria and NIH Guidelines Appendix G.
IV.Adeno-Associated Virus
Adeno-Associated viruses are in the family of viruses Parvoviridae and are considered to be nonpathogenic to humans even though the virus will integrate in the host genome. AAV serotypes 1-4
are classified by the NIH as Risk Group 1. Work with AAV serotypes 1-4 can be conducted at BSL
1. Work with other AAV serotypes will be at a minimum BSL 2 facility and work practices unless
investigator petitions the containment level be downgraded. Using AAV in conjunction with
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Adenovirus or other helper viruses will be at a minimum BSL 2 facility PLUS precautions specific to
work with adenovirus (BSL-2 + Adeno) as described in Section V of this document. Requirements
for BSL 2 facilities and work practices are indicated in BMBL Section IV – Laboratory Biosafety
Level Criteria and NIH Guidelines Appendix G.
V.Laboratory Practices, Containment Equipment (BSL 2 + Adeno)
In addition to complying with the corresponding facility and work practice containment as
specified in the BMBL Section IV – Laboratory Biosafety Level Criteria and if applicable, NIH
Guidelines, work practices and containment for Adenovirus, Adenoviral vectors and AAV
work designated at BSL 2 + Adeno requires the following:
 All vacuum lines must be fitted with a HEPA filter (e.g. “Vacushield J” in-line hydrophobic
filter, Product #4402 from Gelman Sciences).
 No work with Adenovirus is permitted on the open bench. A Biosafety Cabinet must be
used for all manipulations including (but not limited to):
o pipetting
o harvesting infected cells for RNA/DNA/proteins
o loading and opening containers initial delivery of vector in animal hosts
 Reduce risk of exposure by reducing potential of replication competent
adenovirus vector using one of the following methods:
o 1st generation vector – deletion of E1 or E1/E3
o 2nd generation vector – deletion of E1, E3, E4
o 3rd generation vector – all viral gene deleted; only essential cis-acting sequences
retained
 BSL 2 + Adeno includes the use of the following personal protective equipment to
reduce the potential for mucosal exposure, splash to the face, and exposure of
hands:
o Gloves
o Wrap around outer clothing when introducing vector into animals or performing
necropsies. Labcoats are adequate for tissue culture manipulations.
o Goggles (not to be confused with safety glasses)
o N-95 Respirator, to be used with concentrated titers and highly aerosolizing
procedures outside of the Biological Safety Cabinet (contact EH&S for further
information) or in ASBL 2 Facility.
VI.Animal Research with Adenovirus
When animals are infected with adenovirus/adenoviral vectors and some AAV, an Animal BSL2+Adeno (ABSL-2+Adeno) area must be approved and used for the procedure. Concurrent
approvals are needed from the Institutional Biosafety Committee (IBC) and the Institutional Animal
Care and Use Committee (IACUC). All necropsy must be performed in a necropsy room using
Animal BSL-2 + Adenovirus precautionary practices and procedures. Requirements for ABSL-2
containment facility and work practices are indicated in BMBL Section V. Vertebrate Animal
Biosafety Level Criteria for Vivarium Research Facilities.
Infected animals may excrete adenovirus (especially in the first 72 hours after infection).
Precautions must be taken not to create aerosols when emptying animal waste material and when
washing down cages, or cleaning the room with pressure hoses. It is strongly recommended by the
Institutional Biosafety Committee that the lab personnel be responsible for all animal husbandry
practices during the first 72 hours at ABSL-2+Adeno following infection of the animal. After 72
hours, the animals can be housed in an ABSL-1 facility. Contact EH&S for Standard Operating
Procedure for work with ABLS-2+Adeno.
Special training must be given to all animal husbandry personnel on adenovirus, the hazards
associated with the work, required practices and procedures and proper handling of bedding, cage
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washing, and all other husbandry materials associated with the experiment. Contact EH&S at (619)
594-2865 to schedule a group training.
VII.Precautions
Adenovirus is a pathogen of respiratory and gastrointestinal mucous and eye membranes, and
does not have to be replication-competent to cause corneal and conjunctival damage. Goggles
must be worn when working with the agent/vector.
The replication-defective virus may be complemented in vivo thereby causing the vector to become
replication competent.
Adenovirus (unlike HIV or herpes), is quite stable. After having been extracted with ether, and/or
chloroform, it can still be infective.
Signs and labels must be placed to indicate each area where Adenovirus is used or stored
including, but not limited to, Biosafety cabinets, incubators, refrigerators, laboratory entrance doors,
etc.
VIII.Employee Exposure
1. Eye or gastrointestinal mucous membrane exposure from splash or aerosols – rinse a minimum
of 15 minutes in eye wash or flush area with water and take patient immediately to Sharp ReesStealy La Mesa. Notify prior to seeking medical attention the lab manager/principal investigator,
biosafety officer or Human Resources.
2. Inhalation exposure from aerosols – take patient immediately to Sharp Rees-Stealy La Mesa.
Notify prior to seeking medical attention the lab manager/principal investigator, biosafety officer
or Human Resources.
3. Needlestick and/or Sharps Exposure – to prevent inoculation, take patient immediately to Sharp
Rees-Stealy La Mesa. Notify prior to seeking medical attention the lab manager/principal
investigator, biosafety officer or Human Resources.
SYMPTOMATOLOGY: Acute respiratory illness (cold-like symptoms); pneumonia. Conjunctival
infection (or red eye); corneal inflammation.
Contact Information
Sharp Rees Stealy Medical Centers
Occupational Health Services
5525 Grossmont Center Drive
La Mesa, CA 91942
(619) 644-6600
IX.Decontamination
The most effective germicides (with minimum 15 minute contact time) are:

1% Sodium hypochlorite

2% Glutaraldehyde

0.25% Sodium Dodecyl Sulfate
X.Employee Right-to-Know
It is important that all lab personnel (even those not directly working with the virus) be informed and
aware that Adenovirus is being used in the lab.
As the Principal Investigator your signature below indicates that you agree to comply with these
requirements and will acknowledge and accept responsibility for ensuring compliance including all
individuals who enter or work in your laboratory or collaborate in carrying out your research.
Although you may choose to delegate aspects of the biosafety program in your laboratory to other
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laboratory staff, you are ultimately responsible for all activities occurring in your laboratory. Contact
Environmental Health and Safety, Institutional Biosafety Officer at (619) 594-2865 with questions
related these responsibilities.
PI Name:
Signature:
Date:
Return this form to Graduate and Research Affairs, Division of Research Affairs, Student Services
East, Room 1410, MC 8220, retain a copy for your reference.
I have attended laboratory specific safety training for this standard operating procedure. I have
read and understood this standard operating procedure and I have had my questions answered.
Date
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Attendee Name (Print)
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Signature
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