Detection of antibody IgG to HIV-1 in urine by ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24 as antigen for diagnosis of HIV-1 infection

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

• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 848 KB
• Volume: 8
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860080206

✦ Synopsis

Anti-HIV-1 IgG in urine was detected by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24gagprotein (p24) of HIV-1 as antigen and P-D-galactosidase from fscherichia colias IabeLAnti-HIV-1 IgG in urine was reacted simultaneously with 2,4dinitrophenyl-bovine serum albumin-recombinant p24 conjugate and recombinant p24-j3-D-galactosidase conjugate. The complex formed, consisting of the three components, was trapped onto polystyrene balls coated with affinity-purified (anti-2,4dinitrophenyl group) IgG, eluted with~N-2,4dinitrophenyl-L-lysine, and transferred to polystyrene balls coated with aff inity-purified (anti-human IgG y-chain) IgG. Bound P-Dgalactosidase activity was assayed by fluorometry. This assay was at least 3,000-fold more sensitive than conventional methods. The lowest signal among 49 asymptomatic carriers was 3.1-fold higher than the highest nonspecific signal among 100 seronegative subjects. The sensitivity and specificity were both lOO%.The positivitycould beconfirmed by preincubation of urine samples with excess of the antigen.Thus, this assay would be a powerful tool for detecting IgG antibody to HIV-1 in urine. o 1994wiiey-Liss, Inc.