RRML - PCR coupled with mass-spectrometry for detection of Clostridium difficile virulence markers during the emergence of ribotype 027 in Bucharest area

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Nr. 23(4)/2015 DOI:10.1515/rrlm-2015-0044

Short communication

PCR coupled with mass-spectrometry for detection of Clostridium difficile virulence markers during the emergence of ribotype 027 in Bucharest area

Dragos Florea, Steliana Huhulescu, Alexander Indra, Ioana Badicut, Alexandru Rafila, Dan Otelea, Gabriel Adrian Popescu

Abstract:

In recent years Clostridium difficile infection (CDI) has represented a serious public health issue, mainly due to the global spread of the hypervirulent strain NAP1/027/BI. The purpose of the present study was to evaluate the utility of a PCR coupled with electrospray ionization mass spectrometry (ESI-MS) commercial assay for the detection of C. difficile virulence markers. Non-duplicative C. difficile isolates from patients with CDI diagnosed in a tertiary level hospital from Bucharest were tested for toxin A, toxin B, binary toxin genes and deletion in tcdC gene using PCR/capillary gel electrophoresis and PCR/ESI-MS. The study analysed 45 non-duplicative isolates, 33 strains (73.3%) belonging to ribotype 027. The concordance between PCR/capillary gel electrophoresis and PCR/ESI-MS was 100% for toxin A gene, 97.8% for toxin B gene, 91.1% for binary toxin subunit A gene and 95.6% for binary toxin subunit B gene. The general concordance for the complete panel of markers was 88.9% but was 100% for ribotype 027 isolates. PCR/ESI-MS might be a valid method for the detection of C. difficile virulence markers, including binary toxin.

Keywords: Clostridium difficile, ribotype 027, binary toxin, deletion in tcdC gene, electrospray ionization mass spectrometry

Received: 21.8.2015
Accepted: 25.10.2015
Published: 25.11.2015

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Florea D, Huhulescu S, Indra A, Badicut I, Rafila A, Otelea D, et al. PCR coupled with mass-spectrometry for detection of Clostridium difficile virulence markers during the emergence of ribotype 027 in Bucharest area. Rev Romana Med Lab. 2015;23(4):449-55. DOI:10.1515/rrlm-2015-0044