A multiplex, internally controlled real-time PCR assay for detection of toxigenic Clostridium difficile and identification of hypervirulent strain 027/ST-1

作者：Hoegh A M; Nielsen J B; Lester A; Friis Moller A; Schonning K^*

来源：European Journal of Clinical Microbiology & Infectious Diseases, 2012, 31(6): 1073-1079.

DOI：10.1007/s10096-011-1409-5

The purpose of this study was to validate a multiplex real-time PCR assay capable of detecting toxigenic and simultaneously identifying ribotype 027/ST-1 by targeting the toxin genes and in one reaction and in a separate reaction identifying the Delta 117 deletion in associated with ribotype 027/ST-1. PCR was done prospectively on 704 samples routinely submitted to our department and results were compared to results of toxigenic culture. Sequencing of , multi locus sequence typing (MLST) and PCR ribotyping were done on cultured isolates to confirm the correct identification of the Delta 117 deletion in and ribotype 027/ST-1, respectively. The PCR assay displayed a sensitivity, specificity, PPV and NPV of 99.0%, 97.4%, 87.4% and 99.8%, respectively, compared to toxigenic culture on 665 samples evaluable both by PCR and culture. Sequencing of , ribotyping and MLST of cultured isolates validated the genotyping assay and confirmed the ability of the assay to correctly identify ribotype 027/ST-1 in our current epidemiological setting. We describe the use of a combination of two separate PCR assays for sensitive and specific detection of toxigenic and presumptive identification of C. difficile 027/ST-1.

• 出版日期2012-6