       Document 0735
 DOCN  M9640735
 TI    Immune complex transfer enzyme immunoassay that is more sensitive and
       specific than western blotting for detection of antibody immunoglobulin
       G to human immunodeficiency virus type 1 in serum with recombinant pol
       and gag proteins as antigens.
 DT    9604
 AU    Hashida S; Hashinaka K; Nishikata I; Oka S; Shimada K; Saito A;
       Takamizawa A; Shinagawa H; Yano S; Kojima H; et al; Department of
       Biochemistry, Miyazaki Medical College, Japan.
 SO    Clin Diagn Lab Immunol. 1995 Sep;2(5):535-41. Unique Identifier :
       AIDSLINE MED/96089389
 AB    Antibody immunoglobulin G (IgG) to human immunodeficiency virus type 1
       (HIV-1) in serum was detected by ultrasensitive enzyme immunoassays
       (immune complex transfer enzyme immunoassays) with recombinant reverse
       transcriptase (rRT), p17 (rp17) and p24 (rp24) of HIV-1 as antigens and
       beta-D-galactosidase from Escherichia coli as the label. The immune
       complex, comprising 2,4-dinitrophenyl-bovine serum albumin-recombinant
       protein conjugate, antibody IgG to HIV-1, and recombinant
       protein-beta-D-galactosidase conjugate, was trapped on polystyrene beads
       coated with affinity-purified (anti-2,4-dinitrophenyl group) IgG, eluted
       with epsilon N-2,4-dinitrophenyl-L-lysine, and transferred to
       polystyrene beads coated with affinity-purified (anti-human IgG
       gamma-chain) IgG. Bound beta-D-galactosidase activity was assayed by
       fluorometry. The assays were highly reproducible with no serious serum
       interference, and they were much more sensitive than Western
       immunoblotting for the corresponding antigens. Signals with rRT, rp17,
       and rp24 for asymptomatic carriers were at least 56,000-, 680-, and
       22-fold higher, respectively, than those for seronegative individuals,
       and neither indeterminate nor false-positive results were observed,
       whereas some serum samples were false negative or false positive by
       Western blotting for p17 and/or p24 antigen. In some cases,
       seroconversion was detected earlier than by conventional methods.
       Therefore, these assays are suggested to be more useful than
       conventional methods not only for the confirmation of antibody IgGs to
       RT, p17, and p24 of HIV-1 in serum but also for the detection of
       seroconversion.
 DE    *Antigen-Antibody Complex/BLOOD/CHEMISTRY  Blotting, Western
       Comparative Study  Gene Products, gag/*IMMUNOLOGY  Gene Products,
       pol/*IMMUNOLOGY  Human  HIV Antibodies/ANALYSIS/*IMMUNOLOGY  HIV
       Antigens/*IMMUNOLOGY  HIV Seropositivity/BLOOD/DIAGNOSIS  IgG/ANALYSIS
       Immunoenzyme Techniques/STANDARDS  Sensitivity and Specificity  JOURNAL
       ARTICLE

       SOURCE: National Library of Medicine.  NOTICE: This material may be
       protected by Copyright Law (Title 17, U.S.Code).

