Monoclonal antibody solution hybridization assay for detection of human immunodeficiency virus nucleic acids

Abstract

In this report we describe a novel, nonisotopic hybridization assay for the measurement of viral RNA in biological samples. The assay involved a solution-phase reaction between a biotinylated DNA probe and RNA target sequences. Labeled hybrids were detected in an immunoreaction by using a solid-phase anti-biotin antibody and an enzyme-labeled monoclonal antibody specific for DNA-RNA hybrids. This monoclonal antibody solution hybridization assay was compared with an antigen-capture immunoassay for the detection of human immunodeficiency virus in 436 cell culture samples from 60 seropositive patients. The sensitivity and specificity of the hybridization assay were 93.5 and 94.6%, respectively. Detection of human immunodeficiency virus solely by hybridization in the initial sample but not subsequent samples from seven cultures may reflect detection of virus that was present in the patients' lymphocytes but did not replicate in vitro. Since the assay method is adapatable to the detection of either RNA or DNA, it could provide a means for the detection of a wide range of viral nucleic acids.