title: A chemiluminescent, microparticle-membrane capture immunoassay for the detection of antibody to hepatitis B core antigen.
abstract: A chemiluminescent, microparticle-membrane capture immunoassay (CLIA/MMC) for the detection of antibody to hepatitis B core antigen (anti-HBc) is described. The assay utilizes recombinant hepatitis B core antigen coupled to carboxylated latex microparticles. Human polyclonal IgG anti-HBc labelled with acridinium competes with antibody in the sample for a limited number of binding sites on the solid phase. After a 40 min incubation at 40 degrees C, the reaction mixture is transferred to a glass fiber capture membrane and washed. A chemiluminescent signal is produced by addition of alkaline peroxide and is quantitated on a semi-automated reader as described. The CLIA/MMC assay was compared with standard EIA and RIA procedures (Corzyme and Corab, respectively, Abbott Laboratories, North Chicago, IL). Assay sensitivities were RIA greater than CLIA/MMC greater than EIA. A population of 200 normal blood donors showed nearly identical distributions with the CLIA/MMC and RIA (mean = 11% inhibition, SD = 13% for both), compared with the EIA (mean = 13% inhibition, SD = 15%). With a selected plasma population (n = 307), the CLIA/MMC immunoassay showed an excellent correlation (r = 0.94) with both the EIA and RIA procedures. Association of anti-HBc reactivity near assay cutoffs with antibody to hepatitis B surface antigen suggested relative specificity in the order RIA greater than CLIA/MMC greater than EIA. The CLIA/MMC procedure, which can be readily automated, provides a non-istopic alternative to current EIA testing with performance more nearly equivalent to RIA.
authors.affiliation: Abbott Laboratories, Abbott Park, IL 60064.
authors.forename: J
authors.initials: J
authors.lastname: Wolf-Rogers
authors.forename: J A
authors.initials: JA
authors.lastname: Weare
authors.forename: K
authors.initials: K
authors.lastname: Rice
authors.forename: E F
authors.initials: EF
authors.lastname: Robertson
authors.forename: P
authors.initials: P
authors.lastname: Guidinger
authors.forename: O S
authors.initials: OS
authors.lastname: Khalil
authors.forename: G
authors.initials: G
authors.lastname: Madsen
mesh_terms: D000166:Acridines; D001667:Binding, Competitive / Q000276:immunology; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006512:Hepatitis B Core Antigens / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods*; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements; D008863:Microspheres; D011863:Radioimmunoassay; D011994:Recombinant Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000166:Acridines; D006510:Hepatitis B Antibodies; D006512:Hepatitis B Core Antigens; D011994:Recombinant Proteins