《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (9): 2279-2284.doi: 10.16431/j.cnki.1671-7236.2016.09.009

• 生物技术 • 上一篇    下一篇

猪流行性腹泻病毒和猪传染性胃肠炎病毒一步法双重RT-PCR检测方法的建立与应用

胡鸿惠, 南文金, 黄健强, 吴静波, 彭国良   

  1. 韶关学院, 韶关 512005
  • 收稿日期:2016-03-25 出版日期:2016-09-20 发布日期:2016-09-20
  • 通讯作者: 胡鸿惠 E-mail:beijinggd@163.com
  • 作者简介:胡鸿惠(1982-),女,湖南郴州人,硕士,助理研究员,研究方向:动物生产与疾病研究
  • 基金资助:

    广东省科技计划项目(2013B0203606008、2016A040402038);粤北生猪生产及疫病防控协同创新发展中心平台项目;韶关市科技计划项目(2013CX/N02、2013CX/N06、韶科[2015]72号)

Establishment and Application of a Double RT-PCR Method for Detection of PEDV and TGEV

HU Hong-hui, NAN Wen-jin, HUANG Jian-qiang, WU Jing-bo, PENG Guo-liang   

  1. Shaoguan University, Shaoguan 512005, China
  • Received:2016-03-25 Online:2016-09-20 Published:2016-09-20

摘要:

本研究旨在建立一种用于临床腹泻病例中猪流行性腹泻病毒(PEDV)和猪传染性胃肠炎病毒(TGEV)快速检测的一步法双重RT-PCR方法。根据GenBank中收录的PEDV和TGEV的基因序列,分别设计两对特性引物,经最佳反应条件的优化和选择,结果显示,该一步法双重RT-PCR检测方法能同时特异性扩增PEDV和TGEV,该方法可检测到稀释度1×10-5的疫苗毒。应用本方法检测2015年度实验室收集的疑似样品,PEDV阳性率为100%。本研究建立的方法快速、敏感性高、特异性强,可用于临床PEDV和TGEV的快速检测及流行病学调查。

关键词: 猪流行性腹泻病毒(PEDV); 猪传染性胃肠炎病毒(TGEV); 双重RT-PCR

Abstract:

The study was aimed to establish a rapid one-step duplex RT-PCR detection method,which could be used to identify and diagnose PEDV and TGEV in clinical diarrhea cases.According to the gene sequences of PEDV and TGEV from GenBank,two pairs of specific primes were designed.Through optimizing and selecting of the best reaction conditions,we finally pinpointed the duplex one-step RT-PCR detection method with strong specificity,which could detect 1×10-5 diluent degree of vaccine. Suspected samples,which were collected from different pig farms in 2015,were detected of PEDV with 100% positive rate.The method was rapid,high sensitivity and specificity,which could be used for clinical detection of PEDV and TGEV,and also for epidemiological investigation.

Key words: porcine epidemic diarrhea virus (PEDV); porcine transmissible gastroenteritis virus (TGEV); duplex RT-PCR

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