Preliminary performance evaluation on a combined HCV-antigen/antibody detection reagent

Abstract

Objective To develop and preliminarily evaluate a combined HCV antigen/antibody (HCV-Ag/Ab) detection reagent. Methods Two HCV multi-epitope chimeric antigens and 2 anti-HCV core antigen (HCV-cAg) monoclonal antibodies were prepared. An HCV double antigen sandwich and double antibody sandwich ELISA reagent was developed after the antigens and antibodies were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The combined HCV-Ag/Ab reagent was used to examine 903 blood samples. The samples of inconsistent results with a HCV-Ab diagnostic kit were validated by MP HCV BLOT 3.0 reagent. The chi-square test was used for data analysis. Results The relative molecular masses of the 2 chimeric antigens were both ~45 000, consistent with the theoretical expectations. Both antibodies had the right size and high purity. The specificity of the combined reagent reached 100%. The sensitivities of HCV-Ag and HCV-Ab detections were 104 IU/ml and 0.1 national clinical unit/ml, respectively. There was no statistical significance between the results of combined reagent and HCV-Ab diagnostic kit (χ2=0.016 3, P>0.05). The validation reagent confirmed the combined reagent results of 2 samples with discrepant HCV-Ab detection results. Conclusions The combined HCV-Ag/Ab detection reagent has high specificity and sensitivity. It is suitable for blood screening and clinical auxiliary diagnosis. Key words: Hepatitis C antigens; Hepatitis C antibodies; Enzyme-linked immunosorbent assay; Reagent kits, diagnostic