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

Hashida, Seiichi; Hirota, Kouichi; Hashinaka, Kazuya; Saitoh, Atsushi; Nakata, Atsuo; Shinagawa, Hideo; Oka, Shinichi; Shimada, Kaoru; Mimaya, J; Matsushita, Shuzo; Ishikawa, Eiji

doi:10.1002/jcla.1860070610

Cited by 40 publications

(64 citation statements)

References 20 publications

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“…Nevertheless, they could detect antibodies to RT in all 70 urine specimens of seropositive cases tested with 100% specificity. The same immunoassay systems using p17 or p24 were also developed (20,21). Antibodies to p17 and p24 were detected in all 70 urine specimens of seropositive cases with 100% specificity.…”

Section: Discussionmentioning

confidence: 99%

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Saito

Morimoto

Ohara

et al. 1995

Microbiology and Immunology

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HIV-1 Gag protein precursor p55, and its processed products, p17, p24, and p15 were overproduced in Escherichia coli and purified to near homogeneity. To study the antigenic properties and the potentiality as the diagnostic and prognostic reagents, varying amounts of the purified Gag proteins were dotted onto the polyvinylidene difluoride membrane and reacted with 40 sera of HIV-1-infected individuals (35 AC, 1 ARC, and 4 AIDS patients) and 10 sera of normal healthy donors. p55 reacted with 40 (100%) sera of HIV-1 carriers, while p17, p24, and p15 reacted with 37 (92.5%), 35 (87.5%) and 34 (85%) of the 40 sera of HIV-1 carriers, respectively. On the whole, the reaction of p55 was especially strong and that of p15 was the weakest. p55 showed the strongest reaction among the four Gag proteins with all specimens, and it showed a positive reaction with a carrier serum with which none of the processed Gag proteins showed a positive reaction. Therefore, p55 is the most useful antigen among the four Gag proteins for detection of the Gag antibodies and may even be one of the most useful antigens for the diagnosis of HIV-1 infection.

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Section: Discussionmentioning

confidence: 99%

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Saito

Morimoto

Ohara

et al. 1995

Microbiology and Immunology

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“…Thiol groups introduced into rp17 molecules were reacted with 6-maleimidohexanoyl-2,4-dinitrophenyl-bovine serum albumin and maleimide-␤-D-galactosidase (EC 3.2.1.23) (10).…”

Section: Methodsmentioning

confidence: 99%

“…Recently, ultrasensitive enzyme immunoassays (EIAs) (immune complex transfer EIAs) of antibody IgGs to HIV-1 antigens have been developed using recombinant RT, p24, and p17 antigens (10,12,18). Antibody IgGs were allowed to react simultaneously with 2,4-dinitrophenyl antigens and antigenenzyme conjugates.…”

mentioning

confidence: 99%

Earlier Detection of Human Immunodeficiency Virus Type 1 p24 Antigen and Immunoglobulin G and M Antibodies to p17 Antigen in Seroconversion Serum Panels by Immune Complex Transfer Enzyme Immunoassays

Hashida

Ishikawa

Hashinaka

et al. 2000

Clin Diagn Lab Immunol

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For earlier diagnosis of human immunodeficiency virus type 1 (HIV-1) infection, the sensitivities of immune complex transfer enzyme immunoassays for HIV-1 p24 antigen and antibody immunoglobulin G (IgG) to HIV-1 p17 antigen were improved approximately 25-and 90-fold, respectively, over those of the previous immunoassays by performing solid-phase immunoreactions with shaking and increasing the serum sample volumes, and immune complex transfer enzyme immunoassay of antibody IgM to p17 antigen was also performed in the same way as the improved immunoassay of antibody IgG to p17 antigen. By the improved immunoassays, p24 antigen and antibody IgG to p17 antigen were detected earlier in 32 and 53%, respectively, of the HIV-1 seroconversion serum panels tested than before the improvements, and p24 antigen was detected as early as or earlier than HIV-1 RNA by reverse transcriptase-PCR (RT-PCR) in all of the panels tested. In 4 panels out of 19 tested, antibody IgG to p17 antigen or both antibodies IgG and IgM to p17 antigen were detected earlier than p24 antigen and RNA, although the antibody levels declined slightly before their steep increases usually observed after p24 antigen and RNA. Thus, the window period in diagnosis of HIV-1 infection can be shortened by detection of p24 antigen with the improved immunoassay as much as by detection of RNA with RT-PCR and, in some cases, more by detection of antibodies IgG and IgM to p17 antigen with the improved immunoassays than by detections of p24 antigen with the improved immunoassay and RNA with RT-PCR.The positive rates of immunoglobulin G (IgG) antibodies to human immunodeficiency virus type 1 (HIV-1) antigens in serum samples from HIV-1-infected subjects have been reported by the conventional enzyme-linked immunosorbent assay (ELISA) and Western blotting, and it has been generally accepted that the positive rates are high with gp160, gp41 and reverse transcriptase (RT), or p66 (a subunit of RT) as antigens (98 to 100% in asymptomatic carriers, 86 to 100% in patients with AIDS-related complex [ARC], and 77 to 100% in patients with AIDS) but low with gag proteins as antigens (63 to 92% in asymptomatic carriers, 50 to 97% in patients with ARC, and 21 to 77% in patients with AIDS using p24 as antigen and 41% in asymptomatic carriers, 30% in patients with ARC, and 14% in patients with AIDS using p17 as antigen) (2,4,6,8). In seroconversion serum panels, the earliest positive band seen by Western blotting has been observed with p24 in some cases (26,29,34) and with gp160 in other cases (30, 32, 35) but not with p17 (13).Recently, ultrasensitive enzyme immunoassays (EIAs) (immune complex transfer EIAs) of antibody IgGs to HIV-1 antigens have been developed using recombinant RT, p24, and p17 antigens (10,12,18). Antibody IgGs were allowed to react simultaneously with 2,4-dinitrophenyl antigens and antigenenzyme conjugates. The immune complexes of the three components formed were trapped on (anti-2,4-dinitrophenyl group) IgG-coated solid-phase (first solid phase) and, after wash...

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“…Antibody IgGs to p17, p24, and RT of HIV-1 were measured essentially in the same way as described previously for urine samples (13,18,22). An aliquot (10 pl) of serum was incubated for 3 h with 100 fmol each of 2,4-dinitrophenylbovine serum albumin-recombinant protein conjugate and recombinant protein-P-D-galactosidase conjugate, nonspecific rabbit serum (10 pl), and inactive P-D-galactosidase ( S O pg).…”

Section: Immune Complex Transfer Enzyme Lmmunoassays For Antibody Lggmentioning

confidence: 99%

Earlier diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgGs to p17 and reverse transcriptase in serum with enzyme immunoassay

Hashida¹,

Hashinaka²,

Nishikata³

et al. 1996

J. Clin. Lab. Anal.

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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

Cited by 40 publications

References 20 publications

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Overproduction, Purification, and Diagnostic Use of the Recombinant HIV‐1 Gag Proteins, the Precursor Protein p55 and the Processed Products p17, p24, and p15

Earlier Detection of Human Immunodeficiency Virus Type 1 p24 Antigen and Immunoglobulin G and M Antibodies to p17 Antigen in Seroconversion Serum Panels by Immune Complex Transfer Enzyme Immunoassays

Earlier diagnosis of HIV-1 infection by simultaneous detection of p24 antigen and antibody IgGs to p17 and reverse transcriptase in serum with enzyme immunoassay

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