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Testing characteristics of VIDAS Clostridium difficile Toxin A II, Premier Toxins AB, cytotoxin assay and bacterial culture for the detection of toxigenic strains of C difficile



Testing characteristics of VIDAS Clostridium difficile Toxin A II, Premier Toxins AB, cytotoxin assay and bacterial culture for the detection of toxigenic strains of C difficile



Abstracts of the General Meeting of the American Society for Microbiology 103: C-015



Background: C. difficile causes toxin-mediated disease in hospitalized patients. Diagnosis depends upon the detection of toxin A and/or toxin B produced by pathogenic strains. This study compares the diagnostic performance of 2 toxin EIAs, cytotoxin assay, and bacterial culture. Methods: Stool samples submitted for the detection of C. difficile toxin were tested using 4 methods; VIDAS C. difficile Toxin A II (bioMerieux, Hazelwood, MO), Premier Toxins A&B (Meridian Diagnostics, Cincinnati, OH), cytotoxin assay and bacterial isolation. Cytotoxin production was identified by cytopathic effect on MRC-5 cells after 24-48 hour incubation. All positives were confirmed with toxin B-specific antitoxin (Bartels, Famsworth, CT). Bacterial cultures were incubated under anaerobic conditions for 4 days on C. difficile Selective Agar (BBL, Cockeysville, MD). Yellow colonies were confirmed to be C. difficile using the RapID ANA II system (Remel, Lenexa, KS). Isolates were grown for 48-72 hours in chopped meat broth (CMB). CMB supernatant was then tested using the VIDAS EIA and the cytotoxin assay. Results: Four hundred two specimens were tested by all 4 diagnostic methods with 21 positive cultures also being tested by the CMB methods. Positives from the cytotoxin assays and from CMB testing were defined as true positives. Specimens positive by both EIAs but negative for cytotoxin were also considered positive. Overall, 45 (11.2%) were positive while 357 (88.8%) were negative. The diagnostic sensitivity, specificity, positive and negative predictive values were as follows: Cytotoxin assay 82%, 100%, 100%, and 97.0%; Premier Toxin A&B 60.0%, 94.4%, 13.2%, and 94.9%; VIDAS Toxin A 45.5%, 99.7%, 95.2%, and 93.7%, Bacterial culture 37.8%, 98.9%, 81.0%, and 92.7% respectively. Conclusions: Cytotoxin assay remains the most sensitive and specific assay for the detection of pathogenic C. difficile. Although the Premier EIA produced a significant number of unconfirmed positives, it was overall more sensitive than either the VIDAS EIA or culture, each of which detected less than half of all true positives in this study.

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