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建立快速诊断及定型神经毒素对肉毒中毒的治疗、降低死亡率具有十分重要的意义。用一对Ａ型肉毒梭菌特异的寡核苷酸引物，扩增Ａ型肉毒神经毒素基因轻链区域一段４７２ｂｐ的ＤＮＡ片段，对梭状芽胞杆菌属的１０种６８株菌进行了鉴定，并对ＰＣＲ检测Ａ型肉毒神经毒素基因的灵敏度进行了检查。结果表明，所有２０株Ａ型肉毒梭菌ＰＣＲ扩增均为阳性，其余各型肉毒梭菌及其它各种梭菌均为阴性。扩增产物经限制性内切酶分析与预期的酶切片段一致。用溴化乙锭对扩增产物染色，可从３×１０３个细菌中检测出Ａ型肉毒神经毒素基因。用酚－氯仿抽提Ａ型肉毒梭菌全菌体ＤＮＡ进行ＰＣＲ扩增，可从１０ｐｇ的ＤＮＡ中得到清晰的扩增产物。由此可见，该ＰＣＲ扩增系统用于Ａ型肉毒神经毒素基因的检测具有灵敏度高，特异性强等特点，为Ａ型肉毒梭菌的鉴定及肉毒中毒的快速诊断提供了一种新的手段。
The establishment of rapid diagnosis and stereotypic neurotoxin on the treatment of botulism and reduce mortality is of great significance. Using a pair of Clostridium botulinum type specific oligonucleotide primers, a 472 bp DNA fragment of the light chain of the botulinum neurotoxin type A gene was amplified, and 10 of the 68 strains of Clostridium were identified , And examined the sensitivity of PCR for detection of the botulinum neurotoxin type A gene. The results showed that all 20 strains of Clostridium botulinum type A were positive for PCR amplification, and all the other types of Clostridium botulinum and other kinds of Clostridium were negative. The amplified products were analyzed by restriction endonucleases in accordance with the expected restriction endonucleases. The amplification product was stained with ethidium bromide to detect the type A botulinum neurotoxin gene from 3x103 bacteria. Phenol - chloroform extraction of Clostridium botulinum type A bacterial DNA for PCR amplification, from 10pg of DNA to obtain a clear amplification products. Thus, the PCR amplification system for detection of type A botulinum neurotoxin gene has the characteristics of high sensitivity and specificity, and provides a method for the identification of Clostridium botulinum type A and the rapid diagnosis of botulism New means.