摘要
目的比较分析4种肉毒毒素(botulinum neurotoxins,Bo NT)基因分型方法,为四川省监测和食物中毒中肉毒梭菌(Clostridium botulinum)分型鉴定提供可靠的方法。方法使用本实验室保存的6株肉毒梭菌(包括A、B、E型)验证4种肉毒梭菌PCR基因分型鉴定方法——美国食品药品监督管理局(FDA)多重PCR分型方法、国际标准化组织(ISO)的两种多重PCR分型方法和一种实时荧光PCR分型方法,比较方法间的差异,并初步分析差异原因。结果 3种多重PCR方法均可在一个反应中同时检测A、B、E 3种型别肉毒梭菌。ISO多重PCR方法 1中A型检测虽能获得预期条带,但结果条带不清晰。其余两种多重PCR方法在分型检测肉毒梭菌时,可获得清晰的预期条带。实时荧光PCR分型方法能在多重反应体系中同时检测到不同型的肉毒梭菌,但由于荧光标记相同,要获得分型结果需要分别检测各毒素型别。结论美国FDA多重PCR方法和ISO多重PCR方法 2操作较简单易行,可在四川省肉毒梭菌监测中推荐使用。
Objective To compare four PCR genotyping method for Clostridium botulinum,and provide the reliable method for detection and identification of Clostridium botulinum from surveillance and foodborn poisoning in Sichuan Province. Methods Six strains of C. botulinum types A,B and E were used to compare four PCR genotyping method —one multiplex PCR method was from US FDA,two multiplex PCR method and one real-time PCR method were from ISO,and the differences were preliminarily analyzed. Results Three multiplex PCR method could detect C. botulinum types A,B and E in a single reaction. The expected bands for type A were vague using ISO multiplex PCR method 1,whereas bright expected bands could be obtained in the identification of C. botulinum by the other two multiplex PCR method. Real-time multiplex PCR method could detect different types of C. botulinum simultaneously; however,classification should be carried out separately because fluorescent labels were the same. Conclusion Multiplex PCR method from FDA and multiplex PCR method 2 from ISO were relatively simple and could be recommended for C. botulinum surveillance in Sichuan Province.
出处
《中国食品卫生杂志》
2017年第4期445-449,共5页
Chinese Journal of Food Hygiene
基金
基金项目:食品安全国家标准食品微生物学检验肉毒梭菌及肉毒毒素检验(GB4789.12-XXXX)修订
关键词
肉毒梭菌
食物中毒
PCR
基因分型
食源性致病菌
比较
Clostridium botulinum
foodborne poisoning
polymerase chain reaction
genotyping
foodborne pathogenic bacteria
comparative
