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改良分子信标实时PCR快速检测大肠杆菌O_(157):H_7
Authors
刘小立
刘涛
+10 more
吴平芳
庄志雄
张佳峰
张顺祥
扈庆华
李庆阁
王冰
石晓路
贺连华
郑琳琳
Publication date
30 May 2006
Publisher
Abstract
目的建立改良分子信标实时PCR快速检测大肠杆菌O157H7的快速方法。方法根据GenBank公布O157H7的rfbE保守序列,设计一对引物和改良分子信标探针,用FAM荧光剂标记探针的5’端,建立改良分子信标实时PCR检测O157H7的反应体系,应用于食物中毒快速诊断和食品微生物检测。结果共检测11种细菌,只有大肠杆菌O157H7有荧光信号,其余10种全无荧光信号,且与其他细菌无交叉反应,DNA灵敏度为64fg,菌液灵敏度为59cfu/ml或2cfu/PCR反应体系。应用改良分子信标实时PCR反应体系检测10株O157H7均出现特异的荧光信号,无干扰。对89份食品样品进行检测,5份O157H7实时PCR阳性,其余样品为阴性,检测仅需2h。5份阳性样本,经传统方法培养,有3份检出大肠杆菌O157H7。结论改良分子信标实时PCR检测体系快速、灵敏度高,特异性强,可用于大肠杆菌O157H7食物中毒的快速诊断和食品微生物检测,为食源性疾病的分子流行病学调查提供新的检测手段
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Last time updated on 16/06/2016