The E.Coli derived 70 kDa recombinant protein contains sequences from 4 gene products (proteins) of the hepatitis C virus (HCV) were scanned by using 3 different PCR-based techniques in search of the most immunoreactive regions suitable for the development of a diagnostic test for the detection of anti-HCV in human sera. All PCR fragments were cloned with pGEX4-2T expression vector and expressed in E. coli as chimeric proteins with glutathione S-transferase. The most diagnostically relevant proteins identified in this study were then contructed into one recombinant antigen. The protein contains the HCV nucleocapsid, NS3 genotype 1b, NS4 genotype 1b and 1a, and NS5 genotype 1b and 1a immunodominant regions.