Yan Ju-ying, Miao Zi-ping, Lü Hua-kun, Zhou Jia-yue, Chen Yin, Lu Yi-yu, Zhang Yan-jun
Department of Microbiology, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China. Email:
Department of Infectious Disease, Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2013 Dec;34(12):1189-93.
In order to investigate etiology and molecular-epidemiological characteristics of enterovirus associated encephalitis (EAE) in Zhejiang, 2008-2012. Method Cerebrospinal fluid and stool specimens were collected from suspected EAE patients, who were admitted to our hospitals. RD and Hep-2 cell lines were used to isolate enterovirus (EV). Serotypes of these EV isolates were identified through neutralization test by using serotype specific anti-sera. VP1 genes of these isolates were sequenced, compared and used for the construction of phylogenetic tree.
127 (20.6%) human enterovirus (HEV) strains were isolated from 616 samples, which were collected from 610 patients. Serotypes of these EV isolates, including 60 coxsackievirus (CV), and 67 Echovirus (E)appeared to be CVA9, CVB1, CVB3-5, E3, E4, E6, E9, E14, E25 and E30, respectively. Predominant EV serotypes on EAE from 2008 to 2012 were seen as CVB3, CVB5, E6, E30 and E30, respectively. The full length of VP1 genes from different EV isolates was between 834 and 918 nucleotides. The VP1 gene similarities between these isolates and the reference strains were from 76.7% to 85.0% (nucleotides level) and 91.1% to 97.9% (amino acids level). The VP1 genes from E6 serotype isolates appeared most diverged, reaching 20.4% (nucleotides level) and 4.8% (amino acids level). Based on the generated phylogenetic tree, all the EV isolates were fallen on the same branch of HEV-B, and the isolates in the same serotype formed one sub-branch, suggesting there existed geographical and temporal effects. E6 isolates diverged into two branchlets.
EVs from HEV-B were the etiologic agents for EAE in Zhejiang province from 2008 to 2012. All these EV isolates showed 12 serotypes, with predominant isolates varied every year. E30 was determined as the most dominant serotype while serotype E6 diverged into two sub-gene types.
为调查2008 - 2012年浙江省肠道病毒相关性脑炎（EAE）的病因及分子流行病学特征。方法：收集我院收治的疑似EAE患者的脑脊液和粪便标本。采用RD和Hep - 2细胞系分离肠道病毒（EV）。通过使用型特异性抗血清的中和试验鉴定这些EV分离株的血清型。对这些分离株的VP1基因进行测序、比较并用于构建系统发育树。
从610例患者采集的616份样本中分离出127株（20.6%）人肠道病毒（HEV）。这些EV分离株的血清型包括60株柯萨奇病毒（CV）和67株埃可病毒（E），分别为CVA9、CVB1、CVB3 - 5、E3、E4、E6、E9、E14、E25和E30。2008年至2012年EAE的主要EV血清型分别为CVB3、CVB5、E6、E30和E30。不同EV分离株的VP1基因全长在834至918个核苷酸之间。这些分离株与参考菌株的VP1基因相似性在核苷酸水平为76.7%至85.0%，在氨基酸水平为91.1%至97.9%。E6血清型分离株的VP1基因差异最大，核苷酸水平达到20.4%，氨基酸水平达到4.8%。基于生成的系统发育树，所有EV分离株均落在HEV - B的同一分支上，同一血清型的分离株形成一个亚分支，表明存在地理和时间效应。E6分离株分为两个小分支。
2008年至2012年浙江省EAE的病原为HEV - B的EV。所有这些EV分离株显示出12种血清型，每年的主要分离株有所不同。E30被确定为最主要的血清型，而E6血清型分为两个亚基因类型。
