Evaluation of the indirect immunofluorescence assay as a confirmatory test for detecting antibodies to the human immunodeficiency virus.
McHugh Tm, Stites Dp, Casavant Ch, Carlson, J Yee, McVay Pa, MP Busch, Levy Ja
Abstract
McHugh Tm, Stites Dp, Casavant Ch, Carlson, J Yee, McVay Pa, MP Busch, Levy Ja
Abstract
One hundred and eighty-four serum specimens were assayed for antibodies to the human immunodeficiency virus. All specimens were screened with a commercial enzyme immunoassay and confirmed by two indirect immunofluorescence assays. Sera were also assayed by Western blot. Results from sera of 48 healthy heterosexual volunteers were all negative by EIA, IFA, and Western blot. Sera from 50 healthy homosexual men negative by EIA were also negative by IFA and Western blot. Sixty-two patients with persistent generalized lymphadenopathy or newly diagnosed AIDS all were positive by EIA, IFA, and Western blot. Of 24 sera from patients with autoantibodies, with no evidence of AIDS-related diseases, five appeared to be false-positive by EIA, since they were nonreactive by IFA and Western blot. In addition, three other samples contained both autoantibodies and human immunodeficiency virus antibodies. False-positive results were observed in both the EIA and IFA with monoclonal antibodies directed toward the MHC class II antigens DQ and DR. The reactivity of these antibodies could not be distinguished from positive patients' sera, in either EIA or IFA. We conclude that in general indirect immunofluorescence performed well as a confirmatory test after screening by enzyme immunoassay for human immunodeficiency virus antibodies.
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One hundred and eighty-four serum specimens were assayed for antibodies to the human immunodeficiency virus. All specimens were screened with a commercial enzyme immunoassay and confirmed by two indirect immunofluorescence assays. Sera were also assayed by Western blot. Results from sera of 48 healthy heterosexual volunteers were all negative by EIA, IFA, and Western blot. Sera from 50 healthy homosexual men negative by EIA were also negative by IFA and Western blot. Sixty-two patients with persistent generalized lymphadenopathy or newly diagnosed AIDS all were positive by EIA, IFA, and Western blot. Of 24 sera from patients with autoantibodies, with no evidence of AIDS-related diseases, five appeared to be false-positive by EIA, since they were nonreactive by IFA and Western blot. In addition, three other samples contained both autoantibodies and human immunodeficiency virus antibodies. False-positive results were observed in both the EIA and IFA with monoclonal antibodies directed toward the MHC class II antigens DQ and DR. The reactivity of these antibodies could not be distinguished from positive patients' sera, in either EIA or IFA. We conclude that in general indirect immunofluorescence performed well as a confirmatory test after screening by enzyme immunoassay for human immunodeficiency virus antibodies.
Key concepts: Western blot, Immunofluorescence, Immunoassay, Antibody, Autoantibody, Virology, Monoclonal antibody, Virus