Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.
RECOMBINANT IMMUNOBLOT ASSAY FOR THE DETECTION OF ANTIBODIES TO HIV
HIV Detection by Genetic Engineering Methods. Luciw PA and Steimer KS, eds. New York, Marcel Dekker, p. 121-41, 1989.. Unique Identifier : AIDSLINE ICDB/90660258 Truett MA; Chien DY; Calarco TL; DiNello RK; Polito AJ; Chiron Corporation, Emeryville, CA
Abstract:
The Chiron recombinant immunoblot assay (CRIA) was developed to detect antibodies to HIV in human serum. Four recombinant viral antigens were used: p24, the major core protein and gag gene product; p31, a polypeptide from the endonuclease region of the pol gene; and two recombinant envelope polypeptides expressed from the gp41- and gp120-coding regions of the env gene of the AIDS retrovirus (gp41 equivalent and gp120 equivalent antigens). The HIV antigens are coated as bands on a nitrocellulose strip, which also includes two levels of human IgG as positive controls. In tests with 24 patients (pts) with AIDS, 22 pts with AIDS-related complex (ARC), 21 contacts of pts with AIDS or ARC, and 287 blood donor sera that were reactive in one or more commercial human T-lymphotropic virus-III antibody assays, the percentages reactive with CRIA were 100, 100, 100, and 51%, respectively. In addition, changes in antigen response were observed in sequential sera samples from individual pts. The CRIA addresses many of the problems associated with virus-based assays, including low signal with the envelope antigens and the problem of copurification of cross-reacting cellular contaminants. Because recombinant antigens are used, the amount of each antigen can be adjusted to optimize the assay conditions so that even weakly positive samples are detected, thus avoiding false negatives. Each antigen is essentially free of cross-reacting material that may copurify with viral antigens (especially p24); thus, most false-positive samples are eliminated. The strip-assay format permits the addition or substitution of reagents as their relevance for diagnosis of disease progression is determined. For example, reverse transcriptase, the p23 product of the sor region, or the p27 product of the env-lor region of HIV may be useful antigens that could be included on the strip in the future. This assay is simple and rapid, and lends itself well to testing the small numbers of samples that would be encountered in a doctor's office or a diagnostic laboratory. (59 Refs)
Keywords: Acquired Immunodeficiency Syndrome/DIAGNOSIS AIDS-Related Complex/DIAGNOSIS Blood Donors *Cloning, Molecular Enzyme-Linked Immunosorbent Assay Human HIV/*GENETICS/IMMUNOLOGY HIV Antibodies/*ANALYSIS HIV Antigens/GENETICS/IMMUNOLOGY HIV Infections/*DIAGNOSIS HIV Seropositivity/DIAGNOSIS Immunoblotting/*METHODS MONOGRAPH 910228
M9120722
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Important note: Information in this article was accurate in 1991. The state of the art may have changed since the publication date.
