Synthetic peptide-based immunoassay as a supplemental test for HCV infection

Abstract

Introduction: Hepatitis C virus (HCV) is a single strand RNA hepatotrophic virus infecting 170 millions around the world and 20% of Egyptian blood donors. Although there has been significant improvement in the enzyme immunoassays (EIAs) in population screening of HCV infection, the development of a low variability, easy to automate and inexpensive supplemental test to support the current immunoassays was of a major concern to several laboratories. Objectives: In the current study, we embarked on a systematic study to analyze by DNA sequencing several HCV isolates to identify conserved core protein sequences and perform explorative analysis of five synthetic peptides from the core/E1 region in anti-HCV antibody assays. Methods: We designed four synthetic-core specific peptides and an E1-specific peptide. These peptides were used to screen HCV antibodies in sera of 100 HCV positive patients and 100 HCV negative subjects and compared the results with those obtained by the commercial systems based on second and third generation enzyme-linked immunosorbent assays. Results: Our results showed that all peptides detect HCV antibodies in infected sera to varying degrees. The synthetic peptide (a.a. 21–40) of the core protein had 99% sensitivity, 100% specificity and was highly reproducible. Conclusion: The above findings make this core peptide a candidate product for developing a supplemental test for chronic HCV infection in the Egyptian population.