title: Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation.
abstract: Four cell fixation procedures were investigated for their abilities to inactivate human immunodeficiency virus (HIV) and preserve its antigenicity for antibody detection by immunofluorescence in MOLT-4-T4 cells. Air-dried cell smears were fixed in cold acetone, in acetone-methanol (1:1), in acetone-methanol (1:1) followed by 70% ethanol and then methanol, or in paraformaldehyde-acetone. Acetone alone did not inactivate cell-associated HIV, but the other three procedures did. HIV inactivation was achieved by storage of acetone-fixed cells at -70 degrees for 40 days. Antigenicity was measured by immunofluorescence assay titrations of selected human sera, a cerebrospinal fluid, and a gp41 monoclonal antibody. Acetone provided the best fixation as measured by fluorescence intensity and antibody titers. The other fixation methods all yielded weaker fluorescence signals and/or decreased titers. Acetone fixation and storage for 40 days at -70 degrees C provides safe and accurate immunofluorescence assay reagents.
authors.affiliation: Laboratoire de santé publique du Québec, Sainte-Anne-de-Bellevue, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Fauvel
authors.forename: G
authors.initials: G
authors.lastname: Ozanne
mesh_terms: D002460:Cell Line; D005455:Fluorescent Antibody Technique; D006678:HIV / Q000254:growth & development* / Q000276:immunology; D015483:HIV Antibodies / Q000032:analysis*; D015488:HIV Antigens / Q000032:analysis*; D006801:Humans; D014775:Virus Activation
chemical_list: D015483:HIV Antibodies; D015488:HIV Antigens