A novel anti-HIV antibody detection kit, by which anti-HIV-1/2 antibodies were detected based on the counting immunoassay using an auto analyser (PAMIA-50), was evaluated. In an examination using 100 samples each of HIV-1 antibody positive and negative sera, either sensitivity and specificity was 100%. This kit was able to detect all antibodies tested including against HIV-1 subtype A to F, E/F, C/E, B/O, HIV-1 group O, and HIV-2. However, by one of the commercially available kits, one serum containing anti-subtype A antibody was missed. In other comparative studies with the conventional kits using various commercial available panel sera, the same results were obtained. This method was completed within 15 min and was able to examine many samples at one assay. Therefore, this kit is a useful and reliable one if hospitals or institutions had PAMIA-50 in their clinical laboratory.