猪传染性胃肠炎病毒RT-PCR检测方法的建立及应用

Establishment of RT-PCR Method for Detecting Transmissible Gastroenteritis Virus and Its Application

为建立一种操作简便、准确率高的检测方法,对内蒙古地区猪传染性胃肠炎流行情况进行调查,参考GenBank中收录的猪传染性胃肠炎病毒的N基因序列,设计合成了一对特异性引物,针对该基因建立了TGEV的RT-PCR检测方法。应用建立的RT-PCR方法对从内蒙古地区7个盟市采集的204份猪腹泻样品进行了TGEV检测。结果显示,受检的204份样品中有24份样品呈现TGEV阳性,阳性率为11.76%。其中从赤峰市采集的样品TGEV阳性率最高,为20.83%;不同类型样品TGEV的检出率也有差异,其中肛门拭子的检出率最高(15.56%),其次是肠道样品(12.00%),粪便样品的检出率最低(7.87%)。表明所建立的RT-PCR检测方法灵敏度高、特异性强,可用于猪传染性胃肠炎病毒的快速检测。

The research aimed to establish a simple and accurate method for detecting the epidemic situation of transmissible gastroenteritis virus in Inner Mongolia.A pair of specific primers were designed and synthesized according to the sequence of N gene of TGEV published in GenBank.The RT-PCR method was established according to N gene of TGEV.204 diarrhea fecal samples were collected from seven cities of Inner Mongolia,and tested by RT-PCR method.The results showed that 24 samples were TGEV positive,and the positive rate was 11.76%.The positive rate of samples collected from Chifeng city was the highest,up to 20.83%.The pathogen detection rate was different in the types of samples,therein TGEV detection rate of anal swab was the highest with 15.56%,the second was intestine,and the lowest was feces,and their positive rates of TGEV were 12.00% and 7.87%.The result showed that the method was highly sensitive and specific,and can be used for rapid detection of transmissible gastroenteritis virus.