大肠埃希氏菌O157:H7的HDA检测方法的建立

采用自行建立和优化的依赖解旋酶DNA恒温扩增(HDA)检测体系,建立HDA检测方法对大肠埃希氏菌O157:H7进行检测。采用大肠埃希氏菌O157:H7的rfbE基因为目的片段设计特异性引物,建成可快速检测大肠埃希氏菌O157:H7的HDA检测法,进行了特异性和灵敏度试验,并与普通PCR方法进行了比较。结果表明:所建立起的检测法,灵敏度为4.3×103 cfu·mL-1,灵敏度与普通PCR方法相当。大肠埃希氏菌O157:H7的依赖解旋酶DNA恒温扩增检测方法具有普通PCR的特异、灵敏等特点,并且对仪器要求更低,用普通水浴槽即可进行反应。具有广阔的推广前景。

Establishment of the HDA method of detecting Escherichia coli O157:H7

In this paper, a new method, which based on helicase-dependent isothermal DNA amplification (HDA) to detect Escherichia coli O157:H7 of sample, was established. A highly specific set of primers was synthesized to target the rfbE gene of Escherichia coli O157:H7 so as to establish helicase-dependent isothermal DNA amplification method. Specific and sensitivity were tested compared with PCR method. The results showed that the sensitivity of HDA was 4.3×10³ cfu·mL^-1 which was similar to the result of PCR method. Detecting Escherichia coli O157:H7 with HDA is specific and sensitive as well as with PCR method and has lower instrumental requirement .