逆转录-聚合酶链反应在乙型脑炎诊断中的建立

Establishment of reverse transcription semi-nested polymerase chain reaction to the pathogenic diagnosis of JE patients

目的 运用逆转录 -半套式 -聚合酶链反应技术 (RT -semi-nested -PCR) ,建立一种新的早期快速检测乙型脑炎病毒的方法。方法 收集临床诊断的 37例乙脑和 15例非乙脑病人标本 (包括血清、脑脊液 )分别为 6 3份和 30份 ,同时用RT -semi-nested -PCR、IgM捕获法ELISA(MacELISA)进行检测。 结果 血清和脑脊液中可成功检测出JEVRNA ,经1 5 %凝胶电泳 ,出现特异性条带 ,符合预计值 4 0 0bp。结论 RT -semi-nested -PCR对检测JEV感染 ,从实验设计到实验条件 ,都是合理的 ,有较高的敏感性和特异性。可用于乙脑的早期诊断。

Aim To establish a rapid and sensitive method for early diagnosis of Japanese Encephalitis Virus (JEV) infection by RT semi nested PCR Method A total of 63 specimens from the Japanese Encephalitis (JE) patients and 30 specimens from the Non JE patients were examined by RT semi nested PCR for JEV RNA and IgM capture ELISA for specific IgM antibody Result JEV RNA could be detected successfully from the serum and cerebrospinal fluid (CSF) specimens of JEV patients or JEV vaccine As expected,the molecule weight of JEV non code region(NCR) product was as same as 400bp of the standard marker after electrophoresis in 1 5% agarose gel Conclusion Our designed primers,as well as experiment conditions,are optimal,as the high sensitivity and specificity have been shown in our experiments This method can be used in the early diagnosis of JEV infection