散发性中枢神经系统感染患者脑脊液中肠道病毒的检测

摘要

目的 通过对我院散发无菌性脑膜炎和病毒性脑炎成人患者的脑脊液中的肠道病毒检测以期调查本地区就诊患者中散发中枢神经系统肠道病毒感染的基本情况,并对两种检测方法进行了对比.方法 本研究收集了2009-2013年间36例散发无菌性脑膜炎和病毒性脑炎患者的脑脊液.用一种快速、敏感的荧光定量PCR技术检测标本中的各种肠道病毒.同时,使用酶联免疫吸附(ELISA)方法对同组标本进行了埃可病毒,柯萨奇病毒B组1-6型的IgM监测.结果 在所有36例患者的脑脊液标本中有9例经PCR检测为肠道病毒阳性,占25％.其中无菌性脑膜炎、病毒性脑炎和无菌性脑膜脑炎患者中,肠道病毒检出率分别为42.86％,12.50％和11.11％.ELISA法检测同组脑脊液标本仅有2例无菌性脑膜炎患者脑脊液为柯萨奇病毒B组IgM抗体阳性.结论 肠道病毒感染是北京散发病毒性脑炎、无菌性脑膜炎和脑膜脑炎患者的常见病原体,特别是无菌性脑膜炎更多见.荧光定量PCR技术在脑脊液肠道病毒检测方面比ELISA法灵敏度高,但两者联合在肠道病毒的临床分型检测中更有优势.%Objective We detected the enterovirus in the CSF of patients with sporadic aseptic meningitis and viral encephalitis in to investigate the enterovirus infection in this area,and two methods were used in this study.Methods CSF samples of 36 patients with sporadic aseptic meningitis and viral encephalitis were collected during 2009-2013.All the samples were frozen and stored at-80℃ immediately before RNA extraction.Total viral RNA extraction was taken within 3 month after the sample was stored by using an RNA extraction kit according to the method described by the instruction of the kit.Then RNA was reversed into cDNA immediately after the extraction of RNA.cDNA was stored at-80℃.Finaly,we used a rapid and sensitive method for the detection of all members of the enterovirus genus by using real-time PCR.We also detected the IgM of 7enterovirus including Echo virus and Coxsackie virus 1-6 serotype by Enzyme-Linked Immunosorbent Assay(ELISA) method.Results Of all the 36 CSF samples,9 (25％) were positive for enterovirus by PCR method.For patients with aseptic meningitis,viral encephalitis and meningoencephalitis,the proportion were 42.86％,12.50％ and 11.11％.Only 2 samples were positive for Coxsackie virus by ELISA method,and both of them had aseptic meningitis.Conclusion Enterovirus infection is a common cause of sporadic CNS virus infection among patients with viral encephalitis,aseptic meningitis and aseptic meningoencephalitis in Beijing,especially for the patients with aseptic meningitis.Real-time PCR was more sensitive than ELISA,while the better way is to use both of them to detect the serotype of the enterovirus.