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Immunofluorescence assay for human immunodeficiency virus antibody: investigation of cell fixation for virus inactivation and antigen preservation

1989; American Society for Microbiology; Volume: 27; Issue: 8 Linguagem: Inglês

10.1128/jcm.27.8.1810-1813.1989

1098-660X

M. Fauvel, G Ozanne,

Herpesvirus Infections and Treatments

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.