食品科学 ›› 2010, Vol. 31 ›› Issue (12): 193-196.doi: 10.7506/spkx1002-6630-201012044

• 分析检测 • 上一篇    下一篇

食品中大肠杆菌O157的PCR检测与wzy基因的测序分析

李 睿,张忠美,戴诗皎,孙言凤,黄万琪*   

  1. 武汉工业学院生物与制药工程学院
  • 收稿日期:2009-10-21 出版日期:2010-06-15 发布日期:2010-12-29
  • 通讯作者: 黄万琪 E-mail:wanqi2002@yahoo.com.cn
  • 基金资助:

    湖北省自然科学基金重点项目(2009CDA118)

PCR Detection of Escherichia coli O157 in Food and Sequencing of wzy Gene

LI Rui,ZHANG Zhong-mei,DAI Shi-jiao,SUN Yan-feng,HUANG Wan-qi*   

  1. College of Biological and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan 430023, China
  • Received:2009-10-21 Online:2010-06-15 Published:2010-12-29
  • Contact: HUANG Wan-qi E-mail:wanqi2002@yahoo.com.cn

摘要:

大肠杆菌O157:H7 和O157:H- 是重要食品致病菌。从食品中分离得到一株O157:H7 菌株EC5。将该菌株PCR 扩增wzy 基因全长并克隆测序。结果表明,wzy 基因序列与大肠杆菌O157:H7 标准株EDL933 有100% 同源性。从NCBI GenBank 数据库下载O157 菌株wzy 基因序列,针对保守序列设计引物并扩增O157 菌株和非O157 菌株。PCR 结果表明,wzy 基因特异性强,可作为检测O157 菌株的标志基因。本研究为开发大肠杆菌O157 的分子检测技术提供新的研究思路。

关键词: 大肠杆菌O157, PCR 检测, wzy 基因, 测序

Abstract:

A strain of Escherichia coli O157:H7 was isolated from fresh beef and named EC5. EC5 was subjected to PCR amplification and sequencing of full-length wzy gene. There was a 100% similarity between the wzy gene sequence of EC5 and that of Escherichia coli O157:H7 EDL933. Specific primers were designed based on the conserved locus of wzy gene sequences known from NCBI GenBank database for differentiating O157 strains from non-O157 ones by PCR assay. Our findings demonstrate that the wzy gene amplified by PCR is highly specific and provides a promising marker gene for the detection of Escherichia coli O157:H7.

Key words: Escherichia coli O157, PCR detection, wzy gene, sequencing

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