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Journal of Clinical Microbiology, June 2008, p. 1996-2001, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.00032-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Comparison of Real-Time PCR for Detection of the tcdC Gene with Four Toxin Immunoassays and Culture in Diagnosis of Clostridium difficile Infection{triangledown}

Lynne M. Sloan, Brian J. Duresko, Daniel R. Gustafson, and Jon E. Rosenblatt*

Division of Clinical Microbiology, Mayo Clinic, Rochester, Minnesota

Received 7 January 2008/ Returned for modification 25 February 2008/ Accepted 11 April 2008

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.

* Corresponding author. Mailing address: The Mayo Clinic, 200 First St. SW, Rochester, MN 55906. Phone: (507) 284-3050. Fax: (507) 284-4272. E-mail: rosenblatt.jon{at}mayo.edu

{triangledown} Published ahead of print on 23 April 2008.

Journal of Clinical Microbiology, June 2008, p. 1996-2001, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.00032-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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