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目的Ｅ型肉毒中毒是人类肉毒中毒的主要型别之一，本试验旨在建立用于Ｅ型肉毒梭菌鉴定的ＰＣＲ方法。方法用人工合成的寡核苷酸引物扩增Ｅ型肉毒神经毒素基因的一段３６８ｂｐ的ＤＮＡ片段，快速检测Ｅ型肉毒神经毒素基因，对梭状芽胞杆菌属的４０株保藏菌株及３３份土壤标本进行了鉴定，用Ｅ型肉毒梭菌ＣＭＣＣ（Ｂ）６４５０１对其灵敏度进行了检查。结果从所有Ｅ型神经毒素原性菌株或标本均能扩增出目的片段，且能用特定的限制性内切酶切成相应的片段。其它菌株均未能扩增出目的片段。灵敏度试验可从３０个菌体扩增出目的片段。结论此方法用于Ｅ型神经毒素原性梭菌的鉴定具有较高的灵敏度及特异性。
Objective E botulism is one of the major types of botulism in humans and this experiment aimed to establish a PCR method for the identification of Clostridium botulinum type E. Methods A 368-bp DNA fragment of the E-botulinum neurotoxin gene was amplified by using synthetic oligonucleotide primers. The E-botulinum neurotoxin gene was rapidly detected. Forty Strain of Clostridia and 33 Soil samples were identified and their sensitivity was examined using Clostridium botulinum type E (CMCC (B) 64501). Results The target fragments were amplified from all E-type neurotoxin strains or samples and cut into the corresponding fragments using specific restriction endonucleases. Other strains failed to amplify the target fragment. Sensitivity test can amplify the target fragment from 30 bacterial cells. Conclusion This method has high sensitivity and specificity for the identification of E - type Clostridium.