Quantitative real-time PCR for detection of neurotoxin genes of Clostridium botulinum types A,B and C in equine samples |
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| Authors: | Amy L. Johnson Susan C. McAdams-Gallagher Raymond W. Sweeney |
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| Affiliation: | 1. School of Civil, Environmental and Architectural Engineering, Korea University, Seoul 136-714, South Korea;2. IT Convergence Materials R&D Group, Korea Institute of Industrial Technology, Chungnam 330-825, South Korea;3. Department of Environmental Engineering, Daegu University, Gyeongbuk 712-714, South Korea;1. Veterinary Specialized Institute Kraljevo, Zicka 34, 36000 Kraljevo, Serbia;2. Faculty of Veterinary Medicine, University of Belgrade, Bulevar oslobodjenja 18, 11000 Belgrade, Serbia;3. Faculty of Agriculture, University of Belgrade, Nemanjina 6, 11080 Belgrade, Serbia;4. Institute for Forage Crops, Globoder, 37000 Krusevac, Serbia |
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| Abstract: | Botulism in horses in the USA is attributed to Clostridium botulinum types A, B or C. In this study, a duplex quantitative real-time PCR (qPCR) for detection of the neurotoxin genes of C. botulinum types A and B, and a singleplex qPCR for detection of the neurotoxin gene of C. botulinum type C, were optimized and validated for equine gastrointestinal, faecal and feed samples. The performance of these assays was evaluated and compared to the standard mouse bioassay (MBA) using 148 well-characterized samples, most of which were acquired from a repository of veterinary diagnostic samples from cases of botulism: 106 samples positive for C. botulinum (25 type A, 27 type B, 28 type C, 1 type D and 25 type E) and 42 negative samples. The sensitivities of the qPCR assays were 89%, 86% and 96% for C. botulinum types A, B and C, respectively. The overall sensitivity of the mouse bioassay for types A, B and C was 81%. The specificities of the qPCR assays were 99–100% and the specificity of the mouse bioassay was 95%. |
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| Keywords: | Botulism Equine Real-time quantitative PCR Mouse bioassay |
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