Diagnosis of hepatitis C virus (HCV) infection using an immunodominant chimeric polyprotein to capture circulating antibodies: reevaluation of the role of HCV in liver disease.
Structural and nonstructural regions of the HCV-encoded polyprotein have been expressed in recombinant yeast, bacteria, or insect cells and used to capture and measure reactive antibodies circulating in different individuals. The putative nucleocapsid protein (C) and nonstructural proteins 3-5 (NS3-NS5) were found to contain the most immunodominant epitopes. The NS3, NS4, and C regions were expressed in yeast in the form of a fused, chimeric polyprotein (C25) and a capture assay for reactive antibody was developed. This anti-C25 assay detects all previously identified HCV-seropositive cases and provides a substantially more sensitive diagnostic for both acute and chronic HCV infections than the current anti-C100-3 (NS4) assay. Anti-C25 was detected more frequently than anti-C100-3 in chronic, transfusion-associated non-A, non-B hepatitis patients from the United States (95% vs. 71%) and Japan (98% vs. 82%), in cryptogenic cirrhosis patients from the United States (62% vs. 28%), and in hepatitis B surface antigen-negative cases of hepatocellular carcinoma from Japan (83% vs. 63%). These data indicate that HCV has a greater role in these liver diseases than was previously thought. In volunteer United States blood donors sampled following the introduction of anti-C100-3 screening, the prevalence of anti-C25 and anti-C100-3 was 0.5% and 0.08%, respectively.
D000956:Antigens, Viral / Q000276:immunology*; D001782:Blood Donors; D001803:Blood Transfusion; D002213:Capsid / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006508:Hepatitis Antibodies / Q000097:blood*; D006526:Hepatitis C / Q000175:diagnosis* / Q000453:epidemiology / Q000276:immunology; D018937:Hepatitis C Antibodies; D006801:Humans; D015995:Prevalence; D011993:Recombinant Fusion Proteins / Q000276:immunology; D014481:United States / Q000453:epidemiology; D014758:Viral Core Proteins / Q000276:immunology*; D017361:Viral Nonstructural Proteins / Q000276:immunology*
D000956:Antigens, Viral; D000939:Epitopes; D006508:Hepatitis Antibodies; D018937:Hepatitis C Antibodies; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D017361:Viral Nonstructural Proteins