botulism; botulinum toxin type C/D; polymerase chain reaction
Abstract :
[en] Botulism is a rare but serious paralytic illness caused by a nerve toxin that is produced by the bacterium Clostridium botulinum. The economic, medical and alimentary consequences can be catastrophic in case of an epizooty. A polymerase chain reaction (PCR)-based assay was developed for the detection of C. botulinum toxigenic strains type C and D in bovine samples. This assay has proved to be less expensive, faster and simpler to use than the mouse bioassay, the current reference method for diagnosis of C. botulinum toxigenic strains. Three pairs of primers were designed, one for global detection of C. botulinum types C and D (primer pair Y), and two strain-specific pairs specifically designed for types C (primer pair VC) and D (primer pair VD). The PCR amplification conditions were optimized and evaluated on 13 bovine and two duck samples that had been previously tested by the mouse bioassay. In order to assess the impact of sample treatment, both DNA extracted from crude samples and three different enrichment broths (TYG, CMM, CMM followed by TYG) were tested. A 100% sensitivity was observed when samples were enriched for 5 days in CMM followed by 1 day in TYG broth. False-negative results were encountered when C. botulinum was screened for in crude samples. These findings indicate that the current PCR is a reliable method for the detection of C. botulinum toxigenic strains type C and D in bovine samples but only after proper enrichment in CMM and TYG broth.
Disciplines :
Veterinary medicine & animal health Public health, health care sciences & services Immunology & infectious disease
Author, co-author :
Prevot, V.
Tweepenninckx, F.
Van Nerom, E.
Linden, Annick ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Santé et pathologies de la faune sauvage
Content, J.
Kimpe, A.
Language :
English
Title :
Optimization of polymerase chain reaction for detection of Clostridium botulinum type C and D in bovine samples
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