Developing a double-antigen sandwich ELISA for effective detection of human hepatitis B core antibody.
A new double-antigen sandwich ELISA for detecting antibody against the human hepatitis B core antigen (anti-HBc) was developed, with recombinant HBc (rHBcAg) immobilized on the solid phase of the plate and a HRP-rHBcAg conjugation for detection. The rHBcAg was expressed in Escherichia coli and purified by a monoclonal antibody (against HBcAg) specific affinity chromatography. This sandwich ELISA could give a semi-quantitative measurement of anti-HBc concentration in the specimen and was 32-256-folds more sensitive than the competitive ELISA. Total of 942 clinical serum samples were tested in parallel by the sandwich ELISA and the commercially competitive ELISA kit. Overall agreement of 98.4% (927 of 942 cases) was obtained. Ten of 15 (67%) discordant specimens reactive by the sandwich assay but negative by the competitive ELISA resulted from the increased sensitivity of the sandwich assay, as other hepatitis B markers present indicated previously or currently being exposed to HBV.
Deng
Li-Juan
LJ
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, China.
Xu
Yan
Y
Huang
Jian
J
D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006509:Hepatitis B / Q000175:diagnosis*; D006510:Hepatitis B Antibodies / Q000097:blood*; D006512:Hepatitis B Core Antigens / Q000276:immunology*; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D012680:Sensitivity and Specificity
D006510:Hepatitis B Antibodies; D006512:Hepatitis B Core Antigens; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins