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首页> 外文期刊>Journal of Clinical Microbiology >Application of PCR to a clinical and environmental investigation of a case of equine botulism.
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Application of PCR to a clinical and environmental investigation of a case of equine botulism.

机译：PCR在马肉毒杆菌症临床和环境研究中的应用。

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PCR for the detection of botulinum neurotoxin gene types A to E was used in the investigation of a case of equine botulism. Samples from a foal diagnosed with toxicoinfectious botulism in 1985 were reanalyzed by PCR and the mouse bioassay in conjunction with an environmental survey. Neurotoxin B was detected by mouse bioassay in culture enrichments of serum, spleen, feces, and intestinal contents. PCR results compared well with mouse bioassay results, detecting type B neurotoxin genes in these samples and also in a liver sample. Other neurotoxin types were not detected by either test. Clostridium botulinum type B was shown to be prevalent in soils collected from the area in which the foal was raised. Four methods were used to test for the presence of botulinum neurotoxin-producing organisms in 66 soil samples taken within a 5-km radius: PCR and agarose gel electrophoresis (types A to E), PCR and an enzyme-linked assay (type B), hybridization of crude alkaline cell lysates with a type B-specific probe, and the mouse bioassay (all types). Fewer soil samples were positive for C. botulinum type B by the mouse bioassay (15%) than by any of the DNA-based detection systems. Hybridization of a type B-specific probe to DNA dot blots (26% of the samples were positive) and PCR-enzyme-linked assay (77% of the samples were positive) were used for the rapid analysis of large numbers of samples, with sensitivity limits of 3 x 10(6) and 3,000 cells, respectively. Conventional detection of PCR products by gel electrophoresis was the most sensitive method (300-cell limit), and in the present environmental survey, neurotoxin B genes only were detected in 94% of the samples.
机译：PCR用于检测A型至E型肉毒杆菌神经毒素基因，用于调查马肉毒杆菌症病例。通过PCR和小鼠生物测定结合环境调查，重新分析了1985年诊断为毒性传染性肉毒杆菌的小马驹的样本。通过小鼠生物测定法检测到血清，脾脏，粪便和肠内容物的培养物中神经毒素B的含量。 PCR结果与小鼠生物测定结果进行了很好的比较，可以检测这些样品以及肝脏样品中的B型神经毒素基因。两种测试均未检测到其他神经毒素类型。 B型肉毒梭菌在从饲养小马驹的地区收集的土壤中普遍存在。使用四种方法测试在5公里半径范围内采集的66个土壤样品中是否存在产生肉毒杆菌神经毒素的生物：PCR和琼脂糖凝胶电泳（A至E型），PCR和酶联测定（B型） ，碱性粗细胞裂解物与B型特异性探针的杂交以及小鼠生物测定（所有类型）。相比于任何基于DNA的检测系统，通过小鼠生物测定法（15％）对B型肉毒梭菌阳性的土壤样本更少。 B型特异性探针与DNA点印迹的杂交（26％的样品为阳性）和PCR酶联测定（77％的样品为阳性）用于快速分析大量样品，灵敏度限制分别为3 x 10（6）和3,000个单元格。常规的通过凝胶电泳检测PCR产物是最灵敏的方法（限300个细胞），在目前的环境调查中，仅在94％的样品中检测到了神经毒素B基因。

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