用于检测马样本中A型、B型和C型肉毒梭菌神经毒素基因的定量实时聚合酶链反应
Quantitative real-time PCR for detection of neurotoxin genes of Clostridium botulinum types A, B and C in equine samples.
作者信息
Johnson Amy L, McAdams-Gallagher Susan C, Sweeney Raymond W
机构信息
Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, PA 19348, USA.
Department of Clinical Studies, New Bolton Center, University of Pennsylvania School of Veterinary Medicine, Kennett Square, PA 19348, USA.
出版信息
Vet J. 2014 Jan;199(1):157-61. doi: 10.1016/j.tvjl.2013.10.023. Epub 2013 Oct 26.
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%.
美国马匹中的肉毒中毒归因于A型、B型或C型肉毒梭菌。在本研究中，针对马的胃肠道、粪便和饲料样本，优化并验证了用于检测A型和B型肉毒梭菌神经毒素基因的双重定量实时PCR（qPCR）以及用于检测C型肉毒梭菌神经毒素基因的单重qPCR。使用148个特征明确的样本评估了这些检测方法的性能，并与标准小鼠生物测定法（MBA）进行比较，其中大部分样本来自肉毒中毒病例的兽医诊断样本库：106个肉毒梭菌阳性样本（25个A型、27个B型, 28个C型、1个D型和25个E型）和42个阴性样本。qPCR检测方法对A型、B型和C型肉毒梭菌的敏感性分别为89%、86%和96%。小鼠生物测定法对A型、B型和C型的总体敏感性为81%。qPCR检测方法的特异性为99 - 100%，小鼠生物测定法的特异性为95%。
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