猪乙型脑炎病毒RT-PCR检测方法的建立

Development of RT-PCR method for detecting swine janpanese encephalitis virus

根据猪乙型脑炎病毒（JEV）基因序列，设计合成了一对引物，以JEV疫苗株为模板，建立了检测JEV的RT—PCR方法。应用该方法对JEV疫苗株RNA进行扩增，获得与预期大小相符，长度为430bp的特异性目的片段；RT-PCR产物测序结果与文献报道的JEV不同毒株的序列同源性达到98％～100％；敏感性测定该RT-PCR可扩增到10Pg的JEV—RNA。结果表明，建立的RT—PCR方法对JEV的检测敏感性高、特异性强。

A pair of primers were designed and synthesized based on the sequences of the JEV genome, and the RT -PCR method to detect the Japanese Encephalitis Virus （JEV） were development. The 430 bp specifical fragment were obtained from the JEV vaccine strains RNA by this RT -PCR. The test of the gene sequence of RT -PCR product achieved to 98%-100% with that of other JEV strains. The sensitivity of RT -PCR reached to 10 pg JEV RNA. These results showed that the RT -PCR method to detect JEV were of better specificity and higher sensibility.