CAJ | 학술논문

目的调查山东省临沂市脑膜炎/脑炎症候群(AMES)病例中人类肠道病毒病原谱的构成,分析主要病原体埃可病毒30型的基因特征.方法采集AMES病例的脑脊液、粪便和(或)咽拭子标本,所有标本经预处理后,接种RD、Hep-2细胞分离病毒,阳性分离物采用血清学和分子生物学方法鉴定型别,与GenBank检索到的其他Echo30毒株进行同源性比较,并构建VP1完整编码区系统进化树研究其遗传进化规律.结果 2010年4-12月,共采集101例病例的222份标本(脑脊液56份,咽拭子94份,粪便72份),其中34例患者的47份标本(脑脊液4份,咽拭子18份,粪便25份)中分离到HEV,11例鉴定为Echo30.通过同源性分析,发现其核苷酸同源性与2003年山东省泰安市、章丘市及江苏省分离株最近(91.2％～95.4％),与原型株差异较大(17.9％～19.6％)；11株临沂分离株之间核苷酸差异为0～10.2％.系统进化树分析显示,临沂分离株属于基因型6的3个不同分支,提示来自3个传播链的Echo30病毒在临沂市共同循环.结论山东省临沂市2010年AMES病例标本的优势分离株为Echo30,与国外分离株属于不同的进化分支,具有不同的基因特征.
목적 조사산동성림기시뇌막염/뇌염증후군(AMES)병례중인류장도병독병원보적구성,분석주요병원체애가병독30형적기인특정.방법 채집AMES병례적뇌척액、분편화(혹)인식자표본,소유표본경예처리후,접충RD、Hep-2세포분리병독,양성분리물채용혈청학화분자생물학방법감정형별,여GenBank검색도적기타Echo30독주진행동원성비교,병구건VP1완정편마구계통진화수연구기유전진화규률.결과 2010년4-12월,공채집101례병례적222빈표본(뇌척액56빈,인식자94빈,분편72빈),기중34례환자적47빈표본(뇌척액4빈,인식자18빈,분편25빈)중분리도HEV,11례감정위Echo30.통과동원성분석,발현기핵감산동원성여2003년산동성태안시、장구시급강소성분리주최근(91.2％～95.4％),여원형주차이교대(17.9％～19.6％)；11주림기분리주지간핵감산차이위0～10.2％.계통진화수분석현시,림기분리주속우기인형6적3개불동분지,제시래자3개전파련적Echo30병독재림기시공동순배.결론 산동성림기시2010년AMES병례표본적우세분리주위Echo30,여국외분리주속우불동적진화분지,구유불동적기인특정.
Objective To investigate the pathogen spectrum constitution of HEV related to acute meningitis/encephalitis syndrome(AMES) and to analyze the genetic characterization of the echovirus 30 (Echo30) isolates.Methods Cerebrospinal fluid,stool,and/or throat swab specimens from 101 AMES cases were collected for virus isolation with RD and HEp-2 cell lines in Linyi city from April to December,2010.After typing by neutralization test,the entire VP1 gene of the isolates were amplified by RT-PCR and sequenced.Homologous comparison was carried out and phylogenetic analysis were performed.Results HEVs were isolated from 47 specimens of 34 patients(4 from cerebrospinal fluid,18 from throut swab and 25 from stool),with 11 Echo30 isolates identified by microneutralization assay and molecular typing method.In alignment of VP1 sequences with global Echo30 isolates,relative high nucleotide homologies(91.2％ -95.4％ ) with Taian,Zhangqiu and Jiangsu isolates and high divergences( 17.9％ - 19.6％ ) with prototype strain were observed.Nucleotide divergences among Linyi isolates were 0 - 10.2％,and 3 lineages were revealed via phylogenetic analysis,reflecting 3 transmission chains of Echo30 co-circulated in Linyi city,2010.Conclusion Echo30 was the predominant serotype of AMES cases in Linyi city,2010.These isolates possessed considerable divergence with Echo30 from other countries.