Sn-PCR在脑脊液中未知肠道病毒鉴定中的应用  

Identification of unknown enteroviruses in cerebrospinal fluid with Sn-PCR

作　　者：卢囡囡 李红[1] 赵生仓 姜双应 于娟[1] 

机构地区：[1]青海省疾病预防控制中心,青海西宁810007

出　　处：《中国病原生物学杂志》2017年第4期325-327,共3页Journal of Pathogen Biology

基　　金：国家科技重大专项传染病监测技术平台项目(No.2012ZX10004-208)

摘　　要：目的利用肠道病毒分子定型试验,对病毒性脑炎患儿的脑脊液标本进行分子型别鉴定。方法提取患儿脑脊液核酸,利用半巢式聚合酶链反应(Sn-PCR)对肠道病毒VP1区进行特异性扩增并进行序列测定,通过BLAST比对确定肠道病毒的类型,采用Mega6.0软件构建基因进化树。结果 BLAST比对显示该病毒是肠道病毒中的柯萨奇病毒B5(Coxsackievirus group B5,CVB5)。从进化树上看,青海分离株04/CVB5/2010/Qinghai/CHN由2009年山东分离株SD2009-067不断演变而来,而与分离株AH-HK19/CHN/2010/CVB5不在同一进化分支,相距甚远,与FJLY/2011096h分离株亲缘性关系较近。结论成功应用Sn-PCR对患儿脑脊液中未知肠道病毒进行鉴定,确定肠道病毒类型是CVB5型,为肠道病毒引起的脑炎疾病的防控提供参考。Objective To identify genotypes of enteroviruses found in cerebrospinal fluid （CSF） from children with vi- ral encephalitis. Methods The VP1 region of enteroviruses was amplified with a semi-nested polymerase chain reaction （Sn-PCR） and sequenced. The genotype of each enterovirus was determined based on BLAST comparison, and a phyloge- netic tree was constructed with Mega 6.0. Results BLAST comparison indicated that the isolated enterovirus was Cox- sackievirus B5 （CVB5）. Based on the phylogenetic tree, the Qinghai isolate 04/CVB5/2010/Qinghai/CHN evolved from SD2009-067, which was close to the FJLY/2011096h isolate but far from AH-HK19/CHN/2010/CVB5. Conclusion Sn-PCR detected enterovirus type CVB5 in CSF from children with viral encephalitis. This finding may provide a reference for prevention and treatment of encephalitis caused by enteroviruses.

关 键 词：肠道病毒 病毒性脑炎 半巢式聚合酶链反应(Sn-PCR) 

分 类 号：R373.2[医药卫生—病原生物学]