肠道病毒ECHO 19型引起无菌性脑炎流行的诊断与遗传性分析

杨秀惠; 严延生; 何爱华; 陈前进; 张月花

Abstract

杨秀惠,严延生,何爱华,陈前进,张月花.肠道病毒ECHO 19型引起无菌性脑炎流行的诊断与遗传性分析[J].Chinese journal of Epidemiology,2006,27(5):375-378

肠道病毒ECHO 19型引起无菌性脑炎流行的诊断与遗传性分析

Molecular Identification and genetic analysis of an echovirus 19-associated epidemic of aseptic encephalitis

Received:December 23, 2004

DOI：

KeyWord: 肠道病毒ECHO19型脑炎,无菌性遗传性分析

English Key Word: Enterovitvs echovirus type 19 Encephalitis, aseptic Phylogenetic analysis

FundProject:福建省科技重大专项基金资助项目(2004YZ01-2)

Author Name	Affiliation
YANG Xiu-hui	Fujian Center for Disease Control and Prevention, Fuzhou 350001, China
YAN Yan-sheng	Fujian Center for Disease Control and Prevention, Fuzhou 350001, China
HE Ai-hua	Fujian Center for Disease Control and Prevention, Fuzhou 350001, China
CHEN Qian-jin	福建省疾病预防控制中心
ZHANG Yue-hua	福建省疾病预防控制中心

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Abstract:

目的明确引起无菌性脑炎流行的病原,了解其遗传特征。方法收集部分病例脑脊液及双份血清标本,同时应用传统的病毒分离及中和鉴定、RT-PCR扩增肠道病毒目的基因片段及序列分析和双份血清中和抗体变化3种方法进行病因学诊断;并进一步针对病原进行基因序列测定和遗传树图构建分析。结果直接从首例及随后送检的18份脑脊液标本中扩增出肠道病毒核酸进行了早期诊断;病毒分离鉴定及双份血清中和抗体检测结果为:用RD和Hep-2两种细胞从30份脑脊液标本中分离出肠道病毒ECHO 19型16株(阳性分离率为53．33％),检测5例病例双份血清ECHO 19 病毒中和抗体水平,其中4例双份血清抗体阳转或呈4倍及以上升高,同时也证实了肠道病毒 ECHO 19为此次脑炎流行的病因;其中5株病毒VP1区全基因序列与已发表的其他ECHO19病毒株进行比对分析,显示:本次分离株间核苷酸序列同源性为98．9％-100．0％,来自于一个共同的祖先, 而与已发表其他毒株间亲缘性较远,差异性为13．0％-22．4％,属于一新的变异福建株。结论此次无菌性脑炎流行的病原为一变异的ECHO 19病毒;运用分子生物学技术不仅可以达到早期诊断目的,而且可以从分子水平提示病毒的遗传特性。

English Abstract:

Objective To ascertain the pathogen of aseptic encephalitis epidemic in Long-Yan city in Fujian, and to find out the genetic characteristics of the virus. Methods Rapid detection of enteroviral RNA by reverse transcription polymerasechain reaction(RT-PCR) was directly carried out in cerebrospinal fluid(CSF) to isolate and identify the viruses from CSF at the same time, and to detect the neutralization antibody in two serum specimens collected in acute and convalescence phase. Nucleotides of VPl region was also analyzed by constructing phylogenetic tree. Results ECHO 19 infection was rapidly diagnosed and sequence analysed by RT-PCR, and then echovirus type 19 from 16 of 30 CSF samples($3.33%)was isolated and detected using RD and Hep-2 cells simultaneity. The titer of ECHO 19 neutralization antibody became positive or increased by 4 times from acute to convalescence phase in 4 of the S patients.Phylogenetic analyses of the VPl genes of these isolates showed that their nucleotides identity were 98. 9 %-100.0% which were different from those ECHO 19 from GeneBank database by 13.0%一22.4%.Conclusion The etiology of the epidemic of aseptic encephalitis was attributed to ECHO 19. The method of molecular identification not only provided rapid diagnosis of enterovirus infections, but also information about the genetic character of the viruses.

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