猪大肠杆菌、沙门菌及产气荚膜梭菌的多重实时荧光定量PCR检测方法的建立及应用

Chinese Journal of Veterinary Science(2021)

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Abstract
分别针对大肠杆菌irp2基因、沙门菌fim Y基因、产气荚膜梭菌plc基因设计特异性引物和TaqMan探针,并对反应条件进行优化,建立了同时检测3种细菌的多重实时荧光定量PCR检测方法.结果 显示,该方法对大肠杆菌、沙门菌、产气荚膜梭菌的质粒标准品的最低检出限分别为9.28×103,3.09×103,4.37×103拷贝/μL,灵敏性较强;仅特异性扩增大肠杆菌、沙门菌、产气荚膜梭菌,对其他细菌无扩增曲线,有较强特异性;组内与组间变异系数均小于5%,有较好重复性.本试验建立的检测方法为大肠杆菌、沙门菌、产气荚膜梭菌的临床快速检测及流行病学调查提供了一种有效手段.
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