DETECTION OF HEPATITIS-C VIRUS-SPECIFIC CORE PROTEIN IN SERUM OF PATIENTS BY A SENSITIVE FLUORESCENCE ENZYME-IMMUNOASSAY (FEIA)

A protein-capture fluorescence enzyme immunoassay (FEIA) was developed using monoclonal antibodies (mAbs) against recombinant hepatitis C vi rus (HCV) core protein, Four hybridoma cell lines (5E3, 5F11, 515S, 10 80S) were established and characterized. These monoclonal antibodies ( mAbs) each had IgG1 and IgG2 isotypes, and recognized major B cell epi topes within the immunodominant nucleoprotein amino terminal subregion , Using mAb 5F11 as the first antibody to the solid phase and beta-D-g alactosidase-conjugated mAb 5E3 as the second antibody to the protein, we established a specific HCV core protein capturing FEIA capable of detecting as little as 20 pg/ml of recombinant HCV core protein, HCV c ore protein in serum was detectable after treatment with 4.0% polyethy leneglycol, 0.5 M NaOH, and 5% Triton X-100. The results of a peptide inhibition assay indicated that this FEIA is specific for HCV RNA posi tive sera, The quantity of HCV core protein detected in serum was sign ificantly correlated to the level of HCV RNA. The detection limit for HCV core proteins was an HCV RNA per titer of similar to 10(4)/ml, Usi ng this FEIA system, the detection ratio of HCV core protein in patien ts with chronic HCV infection was 92.3% (70/76).