Toxin detection from stool specimens is prerequisite for Clostridium difficile-associated diarrhea and colitis. However, in Japan only one toxin detection kit is commercially available, which requires computerized VIDAS fluorescence reader. In this study we evaluated ImmunoCard Toxin A, which is an enzyme immunoassay with a format of individual cassette and needs no special equipment to perform, by comparing with the VIDAS CDA kit. Of 61 stool specimens 12 were positive and 39 were negative by both assays, 7 were VIDAS positive-ImmunoCard negative, 2 were VIDAS invalid-ImmunoCard negative, and 1 was invalid by both assays. Stool specimens which gave inconsistent results between two assays were subjected to cell culture assay to detect toxin B and culture for C. difficile followed by testing of toxin producibility of isolates. Of 7 VIDAS positive-ImmunoCard negative specimens 5 were negative for cell culture assay and toxigenic C. difficile culture and 2 were positive for cell culture assay. By omitting 3 VIDAS invalid specimens and counting 5 specimens, which were cell culture negative but VIDAS positive, as negative and 2, which were cell culture positive and VIDAS positive, as positive, 12 of the 14 VIDAS positive specimens were ImmunoCard positive (sensitivity, 85.7%) and all 44 VIDAS negative were ImmunoCard negative (specificity, 100%). These results indicate that although in comparison with VIDAS CDA, ImmunoCard Toxin A is slightly less sensitive, ImmunoCard is a rapid, simple, and reliable C. difficile toxin A detection kit, which has the advantage of requiring no special equipment to perform and no centrifuge step, as small as 25 muL of a specimen, and as short as approximately 15 min of time to complete the whole testing.