一起肠道病毒脑炎的病原学快速检测分析  

Rapid detection and diagnosis of human enterovirus caused an encephalitis outbreak in Guangdong,China

作　　者：郑焕英[1] 李晖[1] 曾汉日[1] 吴德[1] 孙立梅[1] 梁文佳[1] 肖红[1] 郭雪[1] 刘冷[1] 柯昌文[1] 

机构地区：[1]广东省疾病预防控制中心,广东广州510300

出　　处：《中国病毒病杂志》2013年第1期37-41,共5页Chinese Journal of Viral Diseases

基　　金：国家"十一五"艾滋病和病毒性肝炎等重大传染病防治科技重大专项(2008ZX10004-008)

摘　　要：目的利用分子生物学检测技术,对2012年5月以来发生在广东省粤西地区罗定市的一起病毒性脑炎疫情进行快速的病原体检测和分子型别的鉴定。方法利用人肠道病毒通用荧光定量RT-PCR对病例的32份脑脊液(CSF)标本和7份粪便标本进行人肠道病毒的核酸快速筛查,再利用半巢式聚合酶链反应(Sn-PCR)对人肠道病毒阳性标本的部分VP1区进行扩增,并对扩增产物进行核苷酸序列测定和分析,使用分子定型方法对人肠道病毒进行型别鉴定。结果人肠道病毒通用荧光定量RT-PCR筛查结果显示32份CSF标本中有26份阳性,7份粪便标本全部阳性;阳性率分别为81.3%和100.0%。对33份阳性标本进行Sn-PCR扩增,22份显示有目的条带,测序结果显示15份CSF标本中13份为埃可病毒(ECV)30、2份为ECV9;7份粪便标本中6份为ECV30、1份为ECV9。结论发生在2012年广东省粤西地区罗定市的一起病毒性脑炎疫情是由以ECV30为主的人肠道病毒引起的;荧光定量RT-PCR法与Sn-PCR法结合核苷酸序列测定和分析可以对人肠道病毒脑炎进行快速检测和病原型别鉴定。Objective To rapidly detect and identify the viral encephalitis caused by human enterovirus.Methods Cerebrospinal fluid（CSF）and stool specimens of the viral encephalitis cases were rapidly screened by using real-time quantitative PCR with universal enterovirus primers.The partial VP1 gene fragment from positive specimens was amplified using semi-nested PCR（Sn-PCR）,sequenced and aligned using BLAST with the corresponding subtypes in the NCBI GenBank.Results Out of 32 CSF specimens tested,26（81.3%）were enterovirus positive by real-time PCR,and all 7（100.0%）stool specimens tested were enterovirus positive.Twenty-six out of 33 positive samples were successfully amplified for the target VP1 gene fragment by Sn-PCR.Sequencing analysis showed that 13 of the 15 CSF specimens were ECV30,the other two were ECV9;6 of the 7 stool specimens were ECV30,only one was ECV9.Conclusions The pandemic of viral encephalitis was proved to be an enterovirus outbreak in Luoding city,west Guangdong province of China.Real-time PCR and Sn-PCR are very effective ways for rapid detection and identification of the intestinal viral encephalitis.

关 键 词：肠道病毒 病毒性脑炎 实时荧光定量PCR 半巢式逆转录聚合酶链反应(Sn-PCR) 病原学 

分 类 号：R373.2[医药卫生—病原生物学]