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Validation of a real-time PCR based method for detection of Clostridium botulinum types C, D and their mosaic variants C-D and D-C in a multicenter collaborative trial.

Abstract
Two real-time PCR arrays based on the GeneDisc(®) cycler platform (Pall-GeneDisc Technologies) were evaluated in a multicenter collaborative trial for their capacity to specifically detect and discriminate Clostridium botulinum types C, D and their mosaic variants C-D and D-C that are associated with avian and mammalian botulism. The GeneDisc(®) arrays developed as part of the DG Home funded European project 'AnibioThreat' were highly sensitive and specific when tested on pure isolates and naturally contaminated samples (mostly clinical specimen from avian origin). Results of the multicenter collaborative trial involving eight laboratories in five European Countries (two laboratories in France, Italy and The Netherlands, one laboratory in Denmark and Sweden), using DNA extracts issued from 33 pure isolates and 48 naturally contaminated samples associated with animal botulism cases, demonstrated the robustness of these tests. Results showed a concordance among the eight laboratories of 99.4%-100% for both arrays. The reproducibility of the tests was high with a relative standard deviation ranging from 1.1% to 7.1%. Considering the high level of agreement achieved between the laboratories these PCR arrays constitute robust and suitable tools for rapid detection of C. botulinum types C, D and mosaic types C-D and D-C. These are the first tests for C. botulinum C and D that have been evaluated in a European multicenter collaborative trial.
AuthorsCedric Woudstra, Hanna Skarin, Fabrizio Anniballi, Bruna Auricchio, Dario De Medici, Luca Bano, Ilenia Drigo, Trine Hansen, Charlotta Löfström, Raditijo Hamidjaja, Bart J van Rotterdam, Miriam Koene, Marie-Hélène Bäyon-Auboyer, Jean-Philippe Buffereau, Patrick Fach
JournalAnaerobe (Anaerobe) Vol. 22 Pg. 31-7 (Aug 2013) ISSN: 1095-8274 [Electronic] England
PMID23669132 (Publication Type: Clinical Trial, Journal Article, Multicenter Study, Research Support, Non-U.S. Gov't, Validation Study)
CopyrightCopyright © 2013 Elsevier Ltd. All rights reserved.
Topics
  • Animals
  • Botulism (diagnosis, microbiology)
  • Clostridium botulinum type C (classification, genetics, isolation & purification)
  • Clostridium botulinum type D (classification, genetics, isolation & purification)
  • Europe
  • Humans
  • Real-Time Polymerase Chain Reaction (methods)
  • Reproducibility of Results

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