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Development of Real-time Quantitative PCR Method for Rapid Detection of Enterohemorrhagic Escherichia coli O157∶H7

LI Dan-dan1,XU Yi-gang2,WANG Sui-jia1,GAO Shen-yang3,LI Yi-jing4   

  1. 1.Animal Quarantine Laboratory Inspection & Quarantine Technology Center of Hainan Entry-exit Inspection & Quarantine Bureau,Haikou 570311,China;2.Animal Quarantine Laboratory Inspection & Quarantine Technology Center of Heilongjiang Entry-exit Inspection & Quarantine Bureau,Harbin 150001,China;3.Department of Animal Husbandry & Veterinary Medicine,Liaoning Medical University,Jinzhou 121001,China;4.College of Veterinary Medicine, Northeast Agricultural University,Harbin 150030,China
  • Received:2014-01-07 Online:2014-07-20 Published:2014-08-21

Abstract: A Real-time quantitative PCR method was developed for the rapid detection of enterohemorrhagic Escherichia coli (EHEC) O157∶H7 based on the primers and TaqMan probes which were designed for the conservative domain of rfbE gene of EHEC O157∶H7.The results showed that the sensitivity of Real-time PCR was 7.3 CFU/mL.20 from 310 samples of meat,egg,milk and its products,animal diarrhea materials and artificial contamination samples were positive detected by Real-time PCR assay,which was in accordance with the testing result according to AOAC standard.The results indicated that Real-time PCR assay was a sensitive,rapid and simple tool.

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