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目的建立用于B型肉毒梭菌的分离鉴定方法,为今后快速检测B型肉毒梭菌奠定基础。方法通过革兰氏染色、动物试验、胶体金免疫层析试验对待测食品标本进行检测,确定为肉毒梭菌,用特异性引物进行PCR扩增分型。结果根据菌株特点分离出肉毒梭菌,菌株和标本均能在253 bp位置扩增出目的片段,测序结果与Gen Bank的已知序列对照检索分析,其与已知B型肉毒毒素序列的一致性达到99%以上。结论此方法可用于B型肉毒梭菌的快速分离和鉴定。
Objective To establish a method for the isolation and identification of Clostridium botulinum type B, which will lay the foundation for the rapid detection of Clostridium botulinum type B in the future. Methods Gram stain, animal experiments, colloidal gold immunochromatography test to be tested samples of food to be identified as Clostridium botulinum, with specific primers for PCR amplification typing. Results Clostridium botulinum was isolated according to the characteristics of the strain, and the target fragment was amplified at 253 bp from the strain and the specimen. The sequencing result was compared with the known sequence of Gen Bank. Consistency of more than 99%. Conclusion This method can be used for the rapid isolation and identification of Clostridium botulinum type B.