CAJ | 학술논문

目的为确证2004年浙江省临海市病毒性脑膜炎暴发疫情的病因,分析病原的遗传变异及进化关系.方法采集患者脑脊液样本60份,采用RD和Hep-2细胞同时分离病毒,中和试验法鉴定病毒型别；对分离株VP1和VP4/VP2基因测序,进行同源性与进化分析.结果从60份脑脊液样本中分离到埃柯病毒30型(E30)19株,分离率为31.7％；对4株E30分离株VP1区核苷酸序列测定,其长度均为876个核苷酸(nt),编码292个氨基酸(aa).临海分离株与E30原型株Bastianni在VP1区的nt和aa同源性分别为82.4％～84.1％和93.5％～ 94.2％；4株临海E30株之间nt和aa的同源性分别为87.1％～99.9％和97.9％～ 100.0％.临海分离株分成两类,同类病毒株间的差异很小,而两类病毒株间的差异很大,nt和aa的最大差异率分别为12.9％和2.1％.与临海E30株同源性最高的为2002-2003年浙江E30株.在VP1基因进化树上,临海E30株分别位于G和H基因亚型分支上,其中临海G基因株与2003年浙江、江苏和山东E30株位于同一进化分支,临海H基因株与2002年浙江诸暨株位于同一进化分支.VP4/VP2区进化分析结果与VP1区相似.结论 2004年临海市病毒性脑膜炎暴发疫情由E30G和H不同基因亚型的E30流行株引起；临海E30株与2002-2003年浙江、江苏和山东E30株具有密切的亲缘关系；H基因亚型株推测为新的E30变异株,首先分离于2002年浙江省.
목적 위학증2004년절강성림해시병독성뇌막염폭발역정적병인,분석병원적유전변이급진화관계.방법 채집환자뇌척액양본60빈,채용RD화Hep-2세포동시분리병독,중화시험법감정병독형별；대분리주VP1화VP4/VP2기인측서,진행동원성여진화분석.결과 종60빈뇌척액양본중분리도애가병독30형(E30)19주,분리솔위31.7％；대4주E30분리주VP1구핵감산서렬측정,기장도균위876개핵감산(nt),편마292개안기산(aa).림해분리주여E30원형주Bastianni재VP1구적nt화aa동원성분별위82.4％～84.1％화93.5％～ 94.2％；4주림해E30주지간nt화aa적동원성분별위87.1％～99.9％화97.9％～ 100.0％.림해분리주분성량류,동류병독주간적차이흔소,이량류병독주간적차이흔대,nt화aa적최대차이솔분별위12.9％화2.1％.여림해E30주동원성최고적위2002-2003년절강E30주.재VP1기인진화수상,림해E30주분별위우G화H기인아형분지상,기중림해G기인주여2003년절강、강소화산동E30주위우동일진화분지,림해H기인주여2002년절강제기주위우동일진화분지.VP4/VP2구진화분석결과여VP1구상사.결론 2004년림해시병독성뇌막염폭발역정유E30G화H불동기인아형적E30류행주인기；림해E30주여2002-2003년절강、강소화산동E30주구유밀절적친연관계；H기인아형주추측위신적E30변이주,수선분리우2002년절강성.
Objective In order to confirm the causes of viral meningitis outbreaks in Linhai county,Zhejiang province in 2004,and to analyze the relationship between hereditary variation and evolution of the pathogen.Methods 60 cerebrospinal fluid(CSF)specimens were collected from the suspected patients.Virus strains from the specimens were isolated with RD and Hep-2 cell lines,and identified through neutralization test.VP1 and VP4/VP2 genes of the isolated viruses were sequenced.Both phylogcnetic and homological trees were also constructed.Results 19 Echovirus type 30(E30)strains were isolated from 60 CSFs,in which E30 accounted for 31.7％.All of the complete VP1 genes in 4 sequenced virus isolates of E30 were composed of 876 nt,encoding 292 amino acids(aa).The identity of nucleotide and amino acid in VP1 gene were 82.4％-84.1％ and 93.5％-94.2％ between the 4 Linhai strains and the prototype strain Bastianni of E30,were 87.1％-99.9％ and 97.9％-100.0％ among the 4 virus strains of E30 from Linhai,respectively.The 4 Linhai strains could be classified into two classes.The diversity of nt and aa was minimal in the same class but obvious between the two classes,with the range of diversities as 12.9％ and 2.1％,respectively.The Linhai E30 strains had maximum similarity with the Zhejiang E30 strains in 2002-2003.The 4 Linhai strains of E30 in the phylogenetic tree of the VP1 gene were attributed into two branches of the G and H genotype,respectively.The G branch also included the E30 strains from Zhejiang,Jiangsu and Shangdong in 2003,while the H branch including E30 strains from Zhuji,Zhejiang in 2002.The phylogenetic tree of VP4/VP2 genes was similar to that of VP1 gene.Conclusion The outbreak of viral meningitis in Linhai county in 2004 was caused by the two classes of E30 strains with G and H genotype existed simultaneously.The Linhai E30 strains had maximum genetic relations to the Zhejiang,Jiangsu and Shangdong strains of E30.The H genotype was inferred to be a new variant strain,which was first isolated in Zhejiang province in 2002.