BACKGROUND: The purpose of this study was to evaluate the newly improved third generation enzyme immunoassay kit, LG HCD 3.0TMB (LG Chemical Ltd., Korea) for the detection of antibodies for the hepatitis C virus. METHODS: The 1,068 clinical samples and 3 seroconversion panels were subjected to compare LG HCD 3.0TMB with Ortho HCV 3.0. The discordant clinical samples were confirmed by RT-PCR (Roche Amplicor HCV test) and RIBA (LG HCD confirm, Ortho RIBA HCV 3.0). Reproducibility was estimated using six samples with different anti-HCV levels for each assay. RESULTS: In clinical samples, concordance between LG HCD 3.0TMB and Ortho HCV 3.0 was 98.8% (1,055 of 1,068 tests) in the screening test. After a repeat test of 13 discordant samples, overall concordance was 99.3% (1,061 of 1,068 tests). In the seroconversion panel testing, the LG HCD 3.0TMB detected HCV antibodies earlier than the Ortho HCV 3.0 in one of the three panels. Results of both EIA assays were constant on the reproducibility test. CONCLUSIONS: Based on the good agreement with Ortho HCV 3.0 and superior seroconversion sensitivity, the LG HCD 3.0TMB assay appears to be suitable for detecting HCV antibodies in clinical laboratories.