Development of RT-PCR method for detecting swine janpanese encephalitis virus
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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.
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Key words:
- japanese encephalitis virus /
- RT-PCR /
- detection
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[1] 吴光华,杨佩英,唐家琪.八种重要传染病的防治[M].第1版.北京:人民军医出版社,2001:129-130. [2] 陈伯权,刘琴兰,孙月英,等.单克隆抗体反向被动血凝和反向被动血凝抑制试验在乙脑快速诊断中的应用[J].中华微生物和免疫学杂志,1987,7(1):43-45.
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