<abstract xmlns="http://www.w3.org/1999/xhtml"><p>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.</p></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.

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

1. National Institute for Infectious Diseases “Prof Dr Matei Bals”, Bucharest, Romania

2. Austrian Agency for Health and Food Safety (AGES) Vienna, Austria

3. University of Medicine and Pharmacy “Carol Davila”, Bucharest, Romania

Revista Romana de Medicina de Laborator

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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

In recent years Clostridium difficile infection (CDI) has represented a serious public health issue, mainly due to the global spread of the hypervirulent...

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

Published Online: Dec 30, 2015

Page range: 449 - 456

Received: Aug 21, 2015

Accepted: Oct 25, 2015

DOI: https://doi.org/10.1515/rrlm-2015-0044

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

© 2015 Dragos Florea et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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

Published Online: Dec 30, 2015

Page range: 449 - 456

Received: Aug 21, 2015

Accepted: Oct 25, 2015

DOI: https://doi.org/10.1515/rrlm-2015-0044

KeywordsClostridium difficile, ribotype 027, binary toxin, deletion in tcdC gene, electrospray ionization mass spectrometry

© 2015 Dragos Florea et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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