摘要
设计了1对引物,利用RT-PCR技术检测乙型脑炎病毒(JEV)。从GenBank中查出收录的31株猪乙型脑炎病毒E基因的已知序列,用DNA star软件对这31株猪乙型脑炎病毒E基因进行同源性分析,以确定扩增的靶序列。以这段靶区域为模板,利用Primer 5软件设计了1对引物,用减毒株SA14-14-2建立了检测乙脑病毒的RT-PCR方法,经敏感性,特异性试验测定,证明该方法敏感,特异;该法可检出样品稀释至256倍的鼠脑毒,相当于0.06个TCID50,对4株河北地区JEV分离株进行检测,结果所设计引物对4株病毒均能扩增出预期的片段。
We used RT-PCR to detect Japanese encephalitis virus with the designed primers, comparing with the serological method, the former was faster and more sensitive . The homology of the sequences of E genes of 31 strains Japanese encephalitis viruses in GenBank was analyzed by DNA-STAR system . According to the definite target sequences,a pair of primers were designed by Primer 5 system, and RT-PCR was established to detect JEV by attenuated strain SA14-14-2 . The results showed that this method was more sensitive and more specific. By the RT-PCR , 0.06 TCIDs0 JEV could be detected. Four HeBei isolates of JEV were detected by RT-PCR using this pair of primers,and that all the four isolates could be amplified by RT-PCR.
出处
《动物医学进展》
CSCD
2006年第7期59-62,共4页
Progress In Veterinary Medicine
