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.
Anti-HIV-1 IgG in urine was detected by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24 gag protein (p24) of HIV-1 as antigen and beta-D-galactosidase from Escherichia coli as label. Anti-HIV-1 IgG in urine was reacted simultaneously with 2,4-dinitrophenyl-bovine serum albumin-recombinant p24 conjugate and recombinant p24-beta-D-galactosidase conjugate. 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 epsilon N-2,4-dinitrophenyl-L-lysine, and transferred to polystyrene balls coated with affinity-purified (anti-human IgG gamma-chain) IgG. Bound beta-D-galactosidase 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 100%. The positivity could be confirmed 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.
Hashida
S
S
Department of Biochemistry, Medical College of Miyazaki, Japan.
Hashinaka
K
K
Hirota
K
K
Saitoh
A
A
Nakata
A
A
Shinagawa
H
H
Oka
S
S
Shimada
K
K
Mimaya
J
J
Matsushita
S
S
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D000936:Antigen-Antibody Complex; D002648:Child; D005260:Female; D015483:HIV Antibodies / Q000652:urine*; D016655:HIV Core Protein p24; D015658:HIV Infections / Q000175:diagnosis* / Q000276:immunology; D006679:HIV Seropositivity / Q000175:diagnosis / Q000276:immunology; D015497:HIV-1 / Q000276:immunology*; D015490:HTLV-I Infections / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D007074:Immunoglobulin G / Q000652:urine*; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins / Q000276:immunology; D012680:Sensitivity and Specificity
D000936:Antigen-Antibody Complex; D015483:HIV Antibodies; D016655:HIV Core Protein p24; D007074:Immunoglobulin G; D011994:Recombinant Proteins