Six hundred diarrheal stool specimens were collected from inpatients and outpatients at local university hospitals for the detection of toxigenic Clostridium difficile using three parallel methods, the BD GeneOhm Cdiff assay, the tissue culture cytotoxicity assay, and a commercially available enzyme-linked fluorescence immunoassay (ELFA) (Vidas C. difficile toxin A and B assay; bioMérieux). Toxigenic C. difficile culture was also performed to further clarify discordant results. During a 3-month study period, 58 (9.7%) of the 600 diarrheal samples examined were positive by the BD GeneOhm Cdiff assay, while the Vidas C. difficile toxin A and B assay and the cytotoxicity assay performed directly on stool samples gave 4.7% and 6.3% positivity rates, respectively. In the case of four samples, BD GeneOhm Cdiff assay results were not evaluable at first because of the presence of PCR inhibitors, but upon repeat testing from the frozen lysates, all of these samples proved to be negative. After resolution with toxigenic culture, the cytotoxicity assay proved to be positive in 55 samples (9.2%), while the ELFA was positive in 37 samples (6.2%). Results of culture and repeated cytotoxicity assays emphasized the importance of the culture method, because the use of ELFA or enzyme immunoassay without a culture method may lead to a substantial portion of toxigenic C. difficile strains being missed.Toxin-producing Clostridium difficile strains are important pathogens among patients who are treated with antibiotics or chemotherapeutic agents not only in the hospital environment but also in the community (3, 6, 10). Since the recognition of outbreaks of C. difficile infection (CDI) caused by C. difficile PCR ribotype 027 in Canada, the United States, and several European countries, rapid and accurate diagnosis of CDI is very important to stop the spread of these strains (7,8,19). In addition, the increasing morbidity and mortality rates associated with CDI and the increasing number of recurrences and therapeutic failures also highlight the need for the development of a rapid and reliable detection method for toxigenic C. difficile in diarrheal feces (12).Only a few laboratories routinely use the tissue culture cytotoxicity and toxin neutralization assays for the detection of toxigenic C. difficile in feces, because they are labor-intensive and time-consuming and standardization is very difficult. Due to their rapid turnaround time, enzyme immunoassays (EIAs) that detect toxin A and/or toxin B in stool are used in most laboratories (11,16). To increase the sensitivity of these tests and in some instances to facilitate epidemiological investigations, culture of C. difficile has become essential. In spite of this, most laboratories use a single toxin detection test on feces for detection of toxigenic C. difficile (4). In the last 10 years, in-house PCR and real-time PCR assays have been developed to detect C. difficile toxin genes. These assays have shown very good sensitivity and specificity and short turnaround times (1, 17). Howev...