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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (26): 59-62.

所属专题： 畜牧兽医

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

猪流行性腹泻病毒RT-PCR检测方法建立及初步应用研究

吴学敏 陈如敬 王隆柏 车勇良 刘玉涛 庄向生 严山 周伦江   

  • 收稿日期:2012-06-13 修回日期:2012-06-25 出版日期:2012-09-15 发布日期:2012-09-15
  • 基金资助:

    福建省农科院青年基金

Development and Preliminary Application of a RT-PCR for Detection of Porcine Epidemic Diarrhea Virus

  • Received:2012-06-13 Revised:2012-06-25 Online:2012-09-15 Published:2012-09-15

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摘要：

为建立一种特异、敏感的PEDV的检测方法。根据GenBank中公布的猪流行性腹泻病毒CV777株的M基因序列，设计了一对特异性引物，扩增长度为680 bp的片段。将PCR产物进行测序，与CV777株、SH5株M基因的同源性分别为99.2%和97.6%。试验表明：该方法具有较好的特异性和敏感性，并能检测到PEDV的RNA浓度下限为100 pg/μL。从福建省的3个地区共采集58份哺乳仔猪腹泻样品，并应用此方法进行检测，其阳性检出率为87.9%。

关键词: 多样性, 多样性

Abstract:

In order to establish specific and sensitive detection methods of porcine epidemic diarrhea virus (PEDV). A pair of primer was designed to amplify a product about 680 bp, according to the sequence of the M gene sequence of PEDV-CV777 strain of PEDV in Genbank. The amplified product was sequenced. Then the homology comparisons among the obtained sequence and the M Gene of the CV777 and SH5 stains were performed. The results showed that it shared 99.2% and 97.6% homology with M Gene of the CV777 and SH5, respectively. This method could be used to test PEDV, and the minimum detectable concentration of the PEDV RNA was 100 pg/μL. The method has been used to detect 58 portion specimens of diarrheal swine from three areas named A, B, and C in Fujian province, and the positive tested rate of PEDV was 87.9%.