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

摘要: 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430 bp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10 pg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。

Abstract: 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.