中国畜牧兽医

• 生物技术 • 上一篇    下一篇

福氏志贺菌2型多重PCR检测方法的建立

朱阵,王婧,张继瑜,魏小娟,周绪正,郭肖,刘翠翠   

  1. 中国农业科学院兰州畜牧与兽药研究所兽药重点开放实验室,甘肃兰州 730050
  • 收稿日期:2014-04-02 出版日期:2014-08-20 发布日期:2014-08-22
  • 通讯作者: 张继瑜,男,博士,研究员,博士生导师。Tel:0931-2115278;E-mail:infzjy@sina.com
  • 作者简介:朱阵(1987—),男,河北人,硕士生,研究方向:福氏志贺菌耐药机理。
  • 基金资助:

    国家自然科学基金(31272603、31101836);国家现代农业肉牛牦牛产业技术体系建设专项(CARS-38)。

Establishment of Multiplex PCR for Detecting of Shigella flexneri Serotype 2

ZHU Zhen, WANG Jing, ZHANG Ji-yu, WEI Xiao-juan, ZHOU Xu-zheng, GUO Xiao, LIU Cui-cui   

  1. Key Open Laboratory of New Veterinary Drug Project, Lanzhou Institute of Animal Science and Veterinary Pharmaceutics, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China
  • Received:2014-04-02 Online:2014-08-20 Published:2014-08-22

摘要: 为构建一种快速、高效、特异性强的检测与鉴定福氏志贺菌的多重PCR方法,本试验以不同血清型志贺菌中的特有基因ipaHgtrⅡ、gtrⅩ和R002为扩增目标来设计相对应的引物,在同一PCR体系中对福氏志贺菌进行检测。通过一系列探索及对反应条件的优化调整,最终建立了可在多种细菌中快速检测出福氏志贺菌2型(2a和2b)的方法。结果表明,应用多重PCR可快速、高效、准确的对福氏志贺菌进行验证和鉴别,对于预防实验室菌种污染和志贺氏菌病的临床诊断与治疗有着重要意义。

关键词: 多重PCR; 福氏志贺菌; 特有基因; 引物

Abstract: In order to build a fast, efficient and specific multiplex PCR method for detection and identification of Shigella flexneri, this study used specific genes ipaH, gtrⅡ, gtrⅩ and R002 in different serotypes of Shigella to design the corresponding primers to amplify the target fragment, and in the same PCR system validated the Shigella flexneri. At the same time through a series of exploration and adjustments to optimize the reaction conditions, we established a method to detect Shigella flexneri type 2a and 2b out of a variety of bacteria quickly. The results suggested that the multiplex PCR could identificate the Shigella flexneri quickly, efficiently and accurately, and had great significance for the prevention of laboratory contamination, clinical diagnosis and treatment of shigellosis.

Key words: multiple PCR; Shigella flexneri; specific genes; primers