Detection 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

✍ Scribed by Seiichi Hashida; Kouichi Hirota; Kazuya Hashinaka; Atsushi Saitoh; Atsuo Nakata; Hideo Shinagawa; Shinichi Oka; Kaoru Shimada; Jun-Ichi Mimaya; Shuzo Matsushita; Dr. Eiji Ishikawa

• Publisher: John Wiley and Sons
• Year: 1993
• Tongue: English
• Weight: 943 KB
• Volume: 7
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860070610

✦ Synopsis

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) US- ing recombinant reverse transcriptase (RT) and p17 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 p-D-galactosidase for pl7. 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 y-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 pos- itivity 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.