Hepatitis C virus (HCV) exposure in blood donors is determined serologically by the detection of anti‐HCV antibodies in serum or plasma. However, a “window” period of 30–70 days after exposure exists where specific antibodies to HCV antigens are not detected. The use of nucleic acid testing for the detection of HCV RNA or antigen testing for the detection of HCV core protein have resulted in dramatic reductions in the pre‐seroconversion window period. In this study, an automated HCV core antigen detection test was developed. This magnetic microparticle‐based assay utilizes anti‐HCV core monoclonal antibody to capture antigen present in human serum or plasma. Captured antigen is then detected using an anti‐HCV core monoclonal antibody conjugated with a chemiluminescent compound. The specificity of this assay was established at 99% upon testing a population of normal volunteer blood donors. Sensitivity was determined by testing 16 commercially available HCV seroconversion panels representing genotypes 1a, 1b, 2b, and 3a. In each panel tested, HCV core antigen was detected prior to anti‐HCV antibody, resulting in a reduction of the window period by greater than 23 days on average, and greater than 34 days on panels initially NAT negative. In addition, HCV core antigen was detected in >97% of HCV RNA positive/antibody negative specimens, exhibiting sensitivity nearly equivalent to nucleic acid testing in the pre‐seroconversion window period for the panels examined. J. Med. Virol. 78:1436–1440, 2006. © 2006 Wiley‐Liss, Inc.