In order to improve the laboratory’s ability on antibody-based assays for human immunodeficiency virus (HIV) detection, a total of 268 serum samples which were positively detected by at least one of the current 3 primary antibody-based assays (chemiluminescence, ELISA and colloidal selenium) were subjected to standard western blot assay for HIV detection in the AIDS Confirmation Laboratory in Dongcheng District, Beijing. The results showed that among 268 samples to be identified, 170 were confirmed positive, with a positive rate of 63.43%. The negative rate of 51 confirmed cases was 19.03%. Inconclusive results accounted for 17.54% of screening responses. The results of this study suggest that for potential HIV infected persons, multiple testing with more than one method should be provided to reduce the risk of missed detection.