Affiliation:
1. Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota
Abstract
ABSTRACT
We have developed a rapid real-time PCR method using fluorescence resonance energy transfer probes and the LightCycler (Roche Diagnostics), which will detect the presence of the
tcdC
gene of
Clostridium difficile
in stool samples. Our PCR method also will identify the presence of base pair deletions, one of which (18 bp) has been associated with the “epidemic” toxin-hyperproducing strains. We compared the results of this PCR with those of three
C. difficile
toxin-detecting enzyme immunoassays (EIAs), an EIA for the detection of glutamate dehydrogenase (GDH), and culture of
C. difficile
. A total of 200 stool specimens were studied by the methods under comparison.
C. difficile
was isolated from 49 specimens by culture, and 44 of these were confirmed as containing one of the genes associated with toxin production (“toxigenic culture”). Using toxigenic culture as the “gold standard”, the sensitivities, specificities, and positive and negative predictive values, respectively, of the assays were 48%, 98%, 88%, and 87% for the Premier toxin A and B test; 48%, 99%, 91%, and 87% for the ImmunoCard toxin A & B test; 48%, 84%, 46%, and 85% for the Xpect
C. difficile
toxin A/B test; 32%, 100%, 100%, and 84% for the Triage
C. difficile
panel (for toxin A); and 86%, 97%, 90%, and 96% for the LightCycler PCR. Thus, in comparison to the sensitivity of toxigenic culture, the sensitivities of the toxin immunoassays were unacceptably low, while the LightCycler real-time PCR assay for the detection of the
tcdC
gene of
C. difficile
is sensitive and specific.
Publisher
American Society for Microbiology
Cited by
184 articles.
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