雷高鹏,杨小蓉,黄玉兰,何树森.四川省肉毒梭菌PCR基因分型方法比较研究[J].中国食品卫生杂志,2017,29(4):445-449. 本文二维码信息
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四川省肉毒梭菌PCR基因分型方法比较研究
Comparative study on PCR genotyping methods of Clostridium botulinum
投稿时间:2017-05-12  
DOI:
中文关键词:  肉毒梭菌  食物中毒  PCR  基因分型  食源性致病菌  比较
Key Words:Clostridium botulinum  foodborne poisoning  polymerase chain reaction  genotyping  foodborne pathogenic bacteria  comparative
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(GB 4789.12—XXXX)修订
作者单位E-mail
雷高鹏 四川省疾病预防控制中心,四川 成都 610041 lionleonleo@126.com,heshs@sohu.com 
杨小蓉 四川省疾病预防控制中心,四川 成都 610041  
黄玉兰 四川省疾病预防控制中心,四川 成都 610041  
何树森 四川省疾病预防控制中心,四川 成都 610041 lionleonleo@126.com,heshs@sohu.com 
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中文摘要:
      目的 比较分析4种肉毒毒素(botulinum neurotoxins,BoNT)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。
Abstract:
      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.
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