摘要
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。
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 syn-thesized 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 Es-cherichia coli O157：H7 with HDA is specific and sensitive as well as with PCR method and has lower instrumen-tal requirement.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2011年第2期271-274,共4页
Journal of Anhui Agricultural University
基金
国家质检总局科研项目(2008IK140)
山东出入境检验检疫局科研项目(SK200821)共同资助
作者简介
王建广，男，硕士研究生。E-mail：jguangwang@sina．cn
通讯作者：雷质文，男，高级兽医师。E—mail：leizhw@sohu．com
