title: Development of a confirmatory enzyme-linked immunosorbent assay for HIV-1 antibodies.
abstract: We subcloned six discrete protein-coding regions representing the gag (Kp24 and Kp55), env (Kp41, Kp120N, and Kp120CC), and pol (Kp66/31) gene products of the human immunodeficiency virus type 1 (HIV-1) and expressed them in Escherichia coli as fusion proteins with the first 56 residues of galactokinase. An enzyme-linked immunosorbent assay for confirming the presence of HIV-1 antibodies was developed by coating the six purified antigens on individual wells of a microtiter plate (the HIVAGEN assay). This assay yielded no false-negative results and fewer indeterminate results than the Western blot assay for 143 specimens from patients with acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC). Analysis of 1016 specimens from seronegative donors by the HIVAGEN assay yielded no false-positive results, and the rate of indeterminate results was substantially lower than for the Western blot assay. The HIVAGEN assay is well suited for routine confirmation of the presence of HIV-1 antibodies because it is objective, quantitative, rapid, precise, and readily automatable.
authors.affiliation: SmithKline Bio-Science Laboratories, Van Nuys, CA 91405.
authors.forename: C S
authors.initials: CS
authors.lastname: Chiang
authors.forename: T
authors.initials: T
authors.lastname: Grove
authors.forename: M
authors.initials: M
authors.lastname: Cooper
authors.forename: J
authors.initials: J
authors.lastname: Cuan
authors.forename: A
authors.initials: A
authors.lastname: Kowalski
authors.forename: K
authors.initials: K
authors.lastname: Parcells
authors.forename: M
authors.initials: M
authors.lastname: Tsunokawa
authors.forename: M
authors.initials: M
authors.lastname: Rosenberg
authors.forename: E
authors.initials: E
authors.lastname: Arcuri
authors.forename: S
authors.initials: S
authors.lastname: Franklin
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis* / Q000276:immunology; D015153:Blotting, Western; D004274:DNA, Recombinant; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D015483:HIV Antibodies / Q000032:analysis*; D015488:HIV Antigens / Q000276:immunology; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D014760:Viral Fusion Proteins / Q000235:genetics
chemical_list: D004274:DNA, Recombinant; D015483:HIV Antibodies; D015488:HIV Antigens; D014760:Viral Fusion Proteins