Objective To evaluate the reliability of enzyme immunoassay (EIA) of HSV-1 as antigen for detection of B virus antibodies. Methods B virus antibodies were detected with enzyme immunoassay (EIA,HSV-1antigen),enzyme-linked immunosorbent assay (ELISA, HSV-1antigen), immunobinding assay (DIA ,B virus antigen), immunofluorecence assay (IFA, B virus antigen) and enzyme-linked immunosorbent assay (ELISA, B virus antigen) in macaque monkey sera. The testing results were compared and analysed. Results Consistency of testing results of EIA(HSV-1antigen)with DIA( B virus antigen), IFA (B virus antigen) and ELISA (B virus antigen)were 100%,95.6% and 97.4%, respectively. The negative sera which were confirmed by EIA (HSV-1antigen) were still negative by IFA( B virus antigen). Among 779 negative sera confirmed by EIA (HSV-1antigen), 2.6%(20) was positive by ELISA (B virus antigen). Conclusion There is much consistency of testing results of EIA(HSV-1antigen)with DIA(B virus antigen), IFA(B virus antigen)and ELISA(B virus antigen). Meanwhile, among B virus free animals which were confirmed using HSV-1antigen, very few animals would be positive using B virus antigen.