儿童病毒性脑膜炎、脑炎中肠道病毒感染的分子生物学分型检测及临床研究  

Molecular biology typing and clinical research of enterovirus infection in children with viral meningitis / encephalitis

作　　者：郭华[1] 丁惠[2] 李慧娟[3] 颜卫红[1] 

机构地区：[1]山东省立医院集团东营医院儿科,山东东营257091 [2]山东省立医院集团东营医院手术室,山东东营257091 [3]山东省立医院集团东营医院血液科,山东东营257091

出　　处：《实用临床医药杂志》2016年第1期35-38,共4页Journal of Clinical Medicine in Practice

基　　金：中国高校医学期刊临床专项资金(11321792)

摘　　要：目的探讨儿童病毒性脑膜炎、脑炎中肠道病毒感染的分子生物学分型。方法采用逆转录聚合酶链反应(RT-PCR)及简并引物扩增并测序,利用Genbank核对分型,检测东营地区2008年1月-2012年2月63例临床诊断为病毒性脑膜炎、脑炎患儿脑脊液中的肠道病毒(EV)RNA,并结合临床特点进行分析。结果 63例病毒性脑膜炎、脑炎患儿的脑脊液经RT-PCR检测,EV感染阳性率为74.6%(47/63)。利用通用引物筛选出的阳性标本,并采用简并引物经两次PCR共获得阳性结果 31例(66.0%)。随机抽取23条阳性片段进行克隆测序,测序结果与Gen Bank标准EV毒株进行同源性对比分析,相关序列同源性均在97%以上。肠道病毒引起中枢神经系统感染的临床特征在各个年龄组有所不同,33例急性期EV患儿脑脊液中白细胞数不同程度增高(均数76×106/L)。结论 EV是儿童无菌性脑膜炎、脑炎的最主要的病原体。采用RT-PCR可以快速准确的检测出中枢神经系统感染的肠道病毒;采用分型引物扩增并测序,将EV的VP1部分序列与GENBANK中的EV标准毒株进行对比,根据基因序列的同源性进行EV型别鉴定是一种切实可行的方案。EV引起的病毒性脑膜炎临床症状一般较轻、无特异性,多数患儿CSF中白细胞高于正常范围。Objective To investigate the molecular biological types of enterovirus（ EV） in infected children with viral meningitis or encephalitis. Methods A Total of 63 children who suffered from viral meningitis / encephalitis between Jan. 2008 to Feb. 2012 at Dongying area were recruited and their cerebrospinal fluid specimens were studied by reverse transcription polymerase chain reaction（ RT-PCR） and degeneracy primers and sequenced to verify the Genbank classification. Their clinical features were also analyzed. Results The positive rate of EV infection was 74. 6 %（ 47 /63）,and the positive results processed by twice PCR was 31 cases（ 62. 0%）. A total of 23 positive fragments were randomly selected to conduct the DNA sequencing,with a result of more than 97% by homologous analysis comparison with Gen Bank standard EV strain. The clinical characteristics of central nervous system infections caused by EV were various form different age groups. The white blood cells in cerebrospinal fluid of the 70. 21% cases with acute EV infection were at a high level（ with a mean of 76× 106/ L）. Conclusion EV is the major pathogen of aseptic meningitis / encephalitis in children. The method of RT-PCR can detect enterovirus infection in central nervous system quickly and accurately.It is a feasible scheme that conducting EV type identification according to the gene sequence homology,which comparing the VP1 partial sequence in EV with standard EV in Genbank. The white blood cells in CSF of most cases are higher above normal range.

关 键 词：肠道病毒 病毒性脑膜炎、脑炎 逆转录聚合酶链反应 

分 类 号：R742[医药卫生—神经病学与精神病学]