逆转录-聚合酶链反应方法检测乙型脑炎病人标本  

Detection of Japanese encephalitis virus in samples of JE patients by RT-PCR

作　　者：孙静[1] 陶三菊[1] 陈伯权[1] 刘琴芝[1] 

机构地区：[1]中国预防医学科学院病毒学研究所虫媒病毒研究室,北京100052

出　　处：《中华实验和临床病毒学杂志》2000年第2期184-187,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　逆转录 聚合酶链反应 (RT PCR)检测乙型脑炎 (乙脑 )病人标本方法的建立和评估。方法　建立RT PCR法 ,了解该方法用于乙脑病毒检测的敏感性 ,特异性 ,并用于临床疑似乙脑病人血清及脑脊液 (CSF)标本的检测 ,并与反向被动血凝抑制实验 (RPHI)方法进行比较分析。结果用该RT PCR法检测高顺生株 (高株 )敏感性可达 6 4PFU。共检测临床疑似乙脑病人标本 38份 ,对CSF中乙型脑炎病毒 (JEV)的检测率明显高于RPHI。结论　RT PCR法有助于临床乙脑病人快速诊断 ,敏感性较高 ,特异性可靠 ,RPHI与RT PCR方法的联合使用将大大提高目前乙脑病人的诊断率。Objective To set up and assess the feasibility of RT PCR method for Japanese encephalitis virus (JEV) detection, and use this method to detect clinical specimens.Methods The sensitivity of this RT PCR was measured by plaque formation test, and the specificity of primers was proved by detecting some other flaviviruses. This RT PCR was also compared with PRHI.Results JEV RNA could be detected successfully from viral culturing supernatants and from mouse brain infected by JEV. Its sensitivity of detecting JEV RNA from viral culturing supernatants was 64 PFU. A total of 38 specimens (of which 25 were serum specimens, 13 CSF specimens) from 16 clinically suspectable JE patients were examined by RT PCR and RPHI.Summing up the detection results of RT PCR and RPHI, 14 patients among the 16 suspectable patients were affirmed as Japanese encephalitis patients. This RT PCR is specific to JEV.The sensitivity of joint use of RT PCR and RPHI to detect JEV was 100%, higher than using RPHI alone, and was 7.7% higher than using RT PCR only. RT PCR is more suitable for epidemiological survey than virus isolation.Conclusion This RT PCR for JEV detection is highy specific and sensitive. Toxemia period of JE patient is very short, so grasping the time of blood specimen collection will affect virus detection rate.

关 键 词：聚合酶链反应 乙型脑炎病毒 病毒检测 

分 类 号：R446.5[医药卫生—诊断学]