Fan Qing-ying, Zhou Xiao-lin, Yang He, Tao Ze-xin, Li Yan, Liu Gui-fang, Liu Yao, Song Li-zhi, Zhang Li, Wang Hai-yan, Xu Ai-qiang
Institute of Epidemiology and Health Statistics, School of Public Health Shandong University, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2009 Dec;23(6):455-7.
To identify the pathogen caused an outbreak of aseptic meningitis in Tancheng county of Shandong province in 2008, and to analyze the molecular characterization of VP1 gene of the Coxsackievirus B3(CVB3) isolates.
Stool and cerebrospinal fluid(CSF) specimens were collected from this outbreak for virus isolation with RD and Hep-2 cell. After typing by neutralization test, the VP1 gene of the isolates were amplified by RT-PCR and sequenced. Homologous comparison and phylogenetic analysis were performed.
35 strains of enteviruse were isolated from 22 stools and 120 CSFs(7 from stools and 28 from CSFs), 34 strains identified as CVB3 and 1 as Echovirus 30(ECHO30) by neutralization test. The nucleotide homologies were 90.5%-100.0% in the partial VP1 gene (381 bp) among 34 CVB3 isolates. Homology comparisons indicated that Shandong strains have the identity of 79.5%-81.6% with the CVB3 prototype strain Nancy. 012/2008TC/SD/CHN and 177/2008TC/SD/CHN showed the highest nucleotides homologies (98.2% and 91.0% respectively) with Fuyang19 strain of Anhui province in 2008 in complete VP1 gene. The phylogenetic tree based on complete VP1 genes showed that all the CVB3 correlated with aseptic meningitis in China recently came from the same evolution linkage and formed a monophyletic cluster.
The causative agent of this outbreak of aseptic meningitis was CVB3. CVB3 circulated in China was genetically different from other countries.
确定2008年山东省郯城县无菌性脑膜炎暴发的病原体，并分析柯萨奇病毒B3（CVB3）分离株VP1基因的分子特征。
从此次暴发中采集粪便和脑脊液标本，用RD和Hep-2细胞进行病毒分离。经中和试验分型后，通过RT-PCR扩增分离株的VP1基因并测序。进行同源性比较和系统发育分析。
从22份粪便和120份脑脊液（粪便7份，脑脊液28份）中分离出35株肠道病毒，经中和试验鉴定34株为CVB3，1株为埃可病毒30型（ECHO30）。34株CVB3分离株部分VP1基因（381 bp）的核苷酸同源性为90.5% - 100.0%。同源性比较表明，山东分离株与CVB3原型株Nancy的同源性为79.5% - 81.6%。012/2008TC/SD/CHN和177/2008TC/SD/CHN在完整VP1基因上与2008年安徽省阜阳19株的核苷酸同源性最高（分别为98.2%和91.0%）。基于完整VP1基因构建的系统发育树显示，近期中国所有与无菌性脑膜炎相关的CVB3均来自同一进化分支，形成一个单系簇。
此次无菌性脑膜炎暴发的病原体为CVB3。在中国流行的CVB3在基因上与其他国家不同。
