【作者】 郑焕英； 孙丽梅； 曾汉日； 吴德； 李晖； 梁文佳； 肖红； 郭雪； 刘冷； 柯昌文；

【Author】 Zheng Huanying,Sun Limei,Zeng Hanri,et al.(Guangdong Provincial Center for Disease Control and Provention, Guangzhou 511430)

【机构】 广东省疾病预防控制中心；

【摘要】 目的:利用分子生物学检测技术,对广东省粤西地区罗定市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EV通用荧光定量PCR对病例CSF和粪便样本进行快速筛查,再利用半巢式聚合酶链反应(Sn-PCR)对阳性样本的部分VP1片段进行扩增和测序,将测序结果登录NCBI网站进行Blast比对,进行EV的型别鉴定。结果:肠道病毒通用荧光定量PCR筛查结果显示32份CSF样本中有26份阳性,7份粪便样本全部阳性;阳性率分别为81.3%和100%。33份阳性样本进行Sn-PCR扩增,22份显示有目的条带,测序结果显示15份CSF样本中13份为ECV30、2份为ECV9;7份粪便样本中6份为ECV30、1份为ECV9。结论:广东省粤西地区罗定市的病毒性脑炎疫情是一起以ECV30为主引起的EV脑炎流行;荧光定量PCR与Sn-PCR结合可以对肠道病毒脑炎进行快速诊断和病原型别鉴定。更多还原

【Abstract】 Objective To rapid detect and identify the viral encephalitis caused by enteroviruses (EV) with molecular biological technology. Methods Cerebrospinal fluid(CSF) and stool specimens of the viral encephalitis cases were rapidly screened by using universal EV primer-Real-time fluorescent quantitative PCR. The VP1 genetic partial fragments of the positive specimens were amplified and sequenced with semi-nested PCR (Sn-PCR). After sequencing, the result was aligned by using BLAST with the corresponding subetypes in the NCBI gene bank. Results Out of 32 CSF specimens tested, 26 specimens was EV positive by Real time -PCR(81.3%), and all 7 stool specimens tested were EV positive by this method(100%). 26 out of 33 positive samples appeared the target bands in the Sn-PCR test. DNA sequence assays 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. Conclusion The pandemic of viral encephalitis was proved to be an EV outbreak in Luoding City, Western Guangdong. Combining the Real-time fluorescent quantitative PCR and Sn-PCR can be a very effective way of rapid detection and identification of the intestinal tract virus encephalitis.更多还原