BACKGROUND: Hepatitis C virus (HCV) has been identified as one of the most frequent causative agent of transfusion associated non-A, non-B hepatitis. So, the pretransfusion screening for anti-HCV is very important to exclude the viral carriers from the blood donor population. Enzyme immunoassay (EIA) is the most commonly used screening assay, but its sensitivity and specificity were not satisfactory yet. To overcome these problems, new assay systems for anti-HCV have been developed and become available for routine use. In this study, we evaluated the newly developed third generation EIA, Genedia(R) HCV ELISA 3.0 and passive hemagglutination assay (PHA), Genedia(R) HCV PHA comparing to the previous third generation EIA kits. METHODS: We performed anti-HCV test with randomly selected 501 serum samples. In all serum samples, we obtained the concordant rates among Genedia(R) HCV ELISA 3.0, Enzymum-Test(R) Anti-HCV and Genedia(R) HCV PHA. In 94 patients' sera, we performed the Abbott(R) HCV EIA 3.0 and obtained the concordant rates with other two EIAs and PHA. Also, we performed HCV confirm test (RIBA HCV 2.0 SIA) and HCV RT-PCR in the sera of discordant results in three EIAs and PHA. RESULTS: The concordant rates among three EIA kits were as followed: Genedia(R) and Enzymum-test, 96.6%; Genedia(R) and Abbott, 97.9%; Enzymum-test(R) and PHA, Abbott(R) and PHA, Genedia(R) HCV ELISA 3.0 and PHA were 96.2%, 95.7% and 98.2%, respectively. In the discordant sera among the three EIAs and PHA, HCV RT-PCR was performed. The concordant rates between Genedia(R) and PCR, Abbott(R) and PCR, PHA and PCR were 94.7%, 84.2% and 78.9%, respectively. And the concordant rate between Enzymum-Test(R) and PCR was only 10.5%. CONCLUSIONS: Genedia(R) HCV ELISA 3.0 and Genedia(R) HCV PHA showed better concordance with Abbott(R) than Enzymum-Test(R) and correlated well with the PCR. Also, Genedia(R) HCV PHA showed relatively satisfactory performance for anti-HCV detection.