Open AccessDetection of antibody IgG to HIV‐1 in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens for diagnosis of HIV‐1 infection
Author(s) -
Hashida Seiichi,
Hirota Kouichi,
Hashinaka Kazuya,
Saitoh Atsushi,
Nakata Atsuo,
Shinagawa Hideo,
Oka Shinichi,
Shimada Kaoru,
Mimaya JunIchi,
Matsushita Shuzo,
Ishikawa Eiji
Publication year - 1993
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860070610
Subject(s) - immunoassay , antigen , immune complex , antibody , recombinant dna , microbiology and biotechnology , affinity chromatography , chemistry , bovine serum albumin , urine , immunoglobulin g , horseradish peroxidase , enzyme , virology , biology , chromatography , biochemistry , immunology , gene
Abstract For diagnosis of HIV‐1 infection, attempts were made to detect anti‐HIV‐1 IgG in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant reverse transcriptase (RT) and p 17 as antigens. Anti‐HIV‐1 IgG in urine was reacted simultaneously with 2,4‐dinitrophenyl‐bovine serum albumin‐recombinant protein conjugate and recombinant proteinenzyme conjugate. The enzymes used as labels were horseradish peroxidase for RT and Escherichia coli β‐ D ‐galactosidase for p17. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity‐purified (anti‐2,4‐dinitrophenyl group) IgG, eluted with ∈N‐2,4‐dinitrophenyl‐L‐lysine and transferred to polystyrene balls coated with affinity‐purified (anti‐human IgG γ‐chain) IgG. Finally, bound enzyme activity was assayed by fluorometry. Urine samples were collected from 100 seronegative subjects and 70 seropositive subjects. The sensitivity and specificity were both 100% with unconcentrated urine samples. The positivity was confirmed by preincubation of urine samples with excess of the antigens. The positivity and negativity with one of the two antigens could be confirmed with the other antigen. The positivity with low signals could be confirmed by concentration of urine samples. Detection of anti‐HIV‐1 IgG in urine by the immune complex transfer enzyme immunoassay using different antigens would make diagnosis of HIV‐1 infection possible. © 1993 Wiley‐Liss, Inc.