Towards an international standard for detection and typing botulinum neurotoxin-producing Clostridia types A, B, E and F in food, feed and environmental samples: a European ring trial study to evaluate a real-time PCR assay.

Lucia Fenicia, Patrick Fach, Bart J van Rotterdam, Fabrizio Anniballi, Bo Segerman, Bruna Auricchio, Elisabetta Delibato, Raditijo A Hamidjaja, Pieter Wielinga, Cedric Woudstra, Joakim Agren, Dario De Medici, Rickard Knutsson

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

A real-time PCR method for detection and typing of BoNT-producing Clostridia types A, B, E, and F was developed on the framework of the European Research Project "Biotracer". A primary evaluation was carried out using 104 strains and 17 clinical and food samples linked to botulism cases. Results showed 100% relative accuracy, 100% relative sensitivity, 100% relative specificity, and 100% selectivity (inclusivity on 73 strains and exclusivity on 31 strains) of the real-time PCR against the reference cultural method combined with the standard mouse bioassay. Furthermore, a ring trial study performed at four different European laboratories in Italy, France, the Netherlands, and Sweden was carried out using 47 strains, and 30 clinical and food samples linked to botulism cases. Results showed a concordance of 95.7% among the four laboratories. The reproducibility generated a relative standard deviation in the range of 2.18% to 13.61%. Considering the high level of agreement achieved between the laboratories, this real-time PCR is a suitable method for rapid detection and typing of BoNT-producing Clostridia in clinical, food and environmental samples and thus support the use of it as an international standard method.
Original languageEnglish
JournalInternational Journal of Food Microbiology
Volume145
Issue numberSuppl 1
Pages (from-to)S152-S157
ISSN0168-1605
DOIs
Publication statusPublished - 2011
Externally publishedYes

Keywords

  • Animal Feed
  • Animals
  • Botulinum Toxins
  • Botulism
  • Clostridium botulinum
  • Clostridium botulinum type A
  • Clostridium botulinum type B
  • Clostridium botulinum type E
  • Clostridium botulinum type F
  • Environmental Microbiology
  • Europe
  • Food Microbiology
  • Humans
  • Mice
  • Molecular Typing
  • Polymerase Chain Reaction
  • Sensitivity and Specificity
  • EC 3.4.24.69 Botulinum Toxins

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