General Guidelines for Experimenting with HIV

Karen B. Byers1, Alan Engelman2, Benjamin Fontes3

1 Dana Farber Cancer Institute, Boston, Massachusetts, 2 Harvard Medical School and Dana Farber Cancer Institute, Boston, Massachusetts, 3 Yale University, New Haven, Connecticut
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 12.1
DOI:  10.1002/0471142735.im1201s59
Online Posting Date:  May, 2004
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Performing experiments using human immunodeficiency virus (HIV)‐infected materials represents a potential biological hazard for the investigator. An intensive training program for laboratory personnel must always precede the actual execution of research involving manipulation of HIV. In addition, appropriate sterile tissue culture techniques are absolute prerequisites for producing meaningful experiments and for achieving safe working conditions. In most circumstances more than one investigator uses the same HIV research facility; therefore, careful training in biosafety is mandatory not only for self‐protection, but for the safety of other investigators as well. The first protocol in this unit establishes a general framework for working safely with HIV and the second describes the proper storage of HIV stocks and test supernatants.

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Table of Contents

  • Basic Protocol 1: General Guidelines for Working Safely with HIV
  • Basic Protocol 2: Preservation of Viral Stocks and Infected Cells
  • Commentary
  • Literature Cited
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Basic Protocol 1: General Guidelines for Working Safely with HIV

  • 70% ethanol
  • Bleach
  • Virucidal disinfectants
  • Biohazard door sign
  • Biohazard labels
  • Biosafety cabinet
  • Tabletop centrifuge and microcentrifuge equipped with aerosol‐containment devices
  • 4°C refrigerator
  • −20° and −80°C freezers
  • Sink equipped with foot pedal
  • Cryogenic storage unit: mechanical −140°C freezer or liquid nitrogen cryogenic storage unit set up for vapor storage only
  • Cryogenic gloves
  • Personal protective equipment: Tyvek suits or surgical gowns (legs must be covered), shoe covers, face shields, disposable sleeve covers, gloves (non‐powdered latex and/or nitrile, with a long cuff)
  • Biohazard bags with temperature‐sensitive labels
  • Medical pathological waste (MPW) disposable containers for incineration
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Key References
   Biosafety in Microbiological and Biomedical Laboratories. US Department of Health and Human Services Publication No. (CDC) 93‐8395. US Government Printing Office, Washington, DC.
  Available online at, these guidelines describe biosafety level 3 practices and summarizes the safety information on retroviruses.
   Centers for Disease Control and Prevention, 2001. See above.
  Available online at, this article contains the latest CDC guidelines for post‐exposure prophylaxis against HIV.
   Sattar and Springthorpe, 1991. See above.
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