Laboratory diagnosis of Hepatitis C virus (HCV) is sometimes problematic. A novel immune enzyme assay (HCV-Core Antigen, Abbott Laboratories/Germany) has been recently licensed for detection of HCV antigen in human serum. In this study, we aimed to evaluate diagnostic performance of this new test in comparison with the HCV-RNA quantification and anti-HCV antibody (anti-HCV-Ab) analyses. A total of 152 serum samples of which 112 anti-HCV-Ab positive (range 1.08 to 386.54 s/co) and 40 anti-HCV-Ab negative (<1 s/co) were analyzed with HCV-RNA and HCV-Ag tests. According to HCV-RNA detection, sensitivity and specificity of HCV-Ag test was measured as 96.9 and 100%, respectively and of anti-HCV-Ab were measured as 100 and 60%, orderly. An excellent positive predictive value for HCV-Ag test was detected as 100%, whereas 28.5% for anti-HCV-Ab test. Pearson correlation analysis showed that there was a statistically significant and strong relationship (p < 0.001, R: 0.773) between HCV-Ag and HCV-RNA quantification analysis. The correlation between the two tests showed an exponential trend (R-2: 0.949). These results suggest that HCV-Ag test may be a reliable assay for HCV antigen detection, which is also well-correlated with serum viral load. However, large studies, including different HCV genotypes and with extreme viral quantity, are required to assess analytic potency of this novel kit.