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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 determine the molecular type of cerebrospinal fluid in children with viral encephalitis using enterovirus molecular typing test. Methods The nucleic acid of cerebrospinal fluid in children was extracted. The VP1 region of enterovirus was amplified by semi-nested polymerase chain reaction (SN-PCR) and sequenced. The type of enterovirus was determined by BLAST. Mega6 was used. 0 Software to build a phylogenetic tree. Results The BLAST alignment showed that the virus was Coxsackievirus group B5 (CVB5) in enterovirus. From the evolutionary tree point of view, Qinghai isolate 04 / CVB5 / 2010 / Qinghai / CHN evolved from Shandong isolate SD2009-067 in 2009 and was not in the same evolutionary branch as the isolate AH-HK19 / CHN / 2010 / CVB5. Far apart, and FJLY / 2011096h isolates closer relationship. Conclusion The successful identification of unknown enterovirus in cerebrospinal fluid of children with Sn-PCR successfully confirmed that the type of enterovirus was CVB5 and provided a reference for prevention and control of enterovirus-induced encephalitis disease.