An innovative molecular detection tool for tracking and tracing Clostridium botulinum types A, B, E, F and other botulinum neurotoxin producing Clostridia based on the GeneDisc cycler

P. Fach, L. Fenicia, R. Knutsson, Pieter Wielinga, F. Anniballi, E. Delibato, B. Auricchio, C. Woudstra, J. Ågren, B. Segerman, D. de Medici, B.J. van Rotterdam

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Rapid and specific detection of botulinum neurotoxin (BoNT) producing Clostridia is a priority for public health authorities, in case of both natural and intentional botulism outbreaks. This study reports on the evaluation of a detection system based on the GeneDisc Cycler designed for simultaneously testing the bont/A, bont/B, bont/E and bont/F genes encoding for the botulinum neurotoxins types A, B, E and F. BoNT-producing Clostridia (n=102) and non-BoNT-producing bacteria (n=52) isolated from clinical, food and environmental samples were tested using this macro-array and results were compared to the reference lethality test on mice. The bont genes were correctly detected in all C. botulinum type A, B, E and F strains available, as well as in toxigenic C. baratii type F and toxigenic C. butyricum type E. No cross reactivity was observed with non human-toxigenic bacteria, C. botulinum types C, D and G. The identification of the bont genotype using the macro-array was correlated to toxino-typing of the BoNTs as determined by the mouse bioassay. An “evaluation trial” of the GeneDisc array performed blind in four European laboratories with 77 BoNT-producing Clostridia as well as 10 food and clinical samples showed that the developed macro-array is specific and reliable for identifying BoNT/A-, BoNT/B-, BoNT/E- and BoNT/F-producing clostridial strains and for screening naturally contaminated food and fecal samples. The test is robust, has a low detection limit (c.a. 5 to 50 genome copies in the PCR reaction microwell) and is promising for monitoring BoNT-producing Clostridia in different kinds of samples including food and clinical samples.
Original languageEnglish
JournalInternational Journal of Food Microbiology
Volume145
Pages (from-to)S145-S151
ISSN0168-1605
DOIs
Publication statusPublished - 2011
Externally publishedYes

Keywords

  • Clostridium botulinum
  • Real-time PCR
  • Macro-array

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