CAJ | 학술논문

目的分析云南中缅边境地区新分离蚊媒病毒的分子生物学特征,以分析其遗传进化特征.方法 2010年1-12月在云南省德宏州芒市和瑞丽市监测点每月一次用诱蚊器采集蚊虫,用BHK-21和C6/36细胞分离病毒,阳性分离物用RT-PCR法进行鉴定及序列测定和分析.结果从采获的7属42种69 209只蚊虫中分离到13株流行性乙型脑炎病毒(Japanese encephalitis virus,JEV),4株盖塔病毒(Getah virus,GETV),1株西藏环状病毒(Tibet orbivirus,TIBOV),7株淡色库蚊浓核病毒(Culex pipiens pallens densovirus,CppDNV).JEV E基因进化分析证实,新分离的13株JEV均属基因Ⅰ型,与云南其它地区和邻近省份流行株亲缘关系较近,且抗原和毒力位点未发生明显改变.4株GETVE2基因与云南其它地区分离株同源性高,亲缘关系近.新分离环状病毒的VP1基因与TIBOV亲缘关系最近,而与云南环状病毒(Yunnan orbivirus)存在明显差异.7株CppDNV的NS1基因与以往云南及其他省区流行株具有较高的同源性和亲缘关系.结论首次证实该地区存在基因Ⅰ型JEV、GETV、TIBOV和CppDNV流行,这些流行株均具有较为稳定的遗传特征和地域特征.
목적 분석운남중면변경지구신분리문매병독적분자생물학특정,이분석기유전진화특정.방법 2010년1-12월재운남성덕굉주망시화서려시감측점매월일차용유문기채집문충,용BHK-21화C6/36세포분리병독,양성분리물용RT-PCR법진행감정급서렬측정화분석.결과 종채획적7속42충69 209지문충중분리도13주류행성을형뇌염병독(Japanese encephalitis virus,JEV),4주개탑병독(Getah virus,GETV),1주서장배상병독(Tibet orbivirus,TIBOV),7주담색고문농핵병독(Culex pipiens pallens densovirus,CppDNV).JEV E기인진화분석증실,신분리적13주JEV균속기인Ⅰ형,여운남기타지구화린근성빈류행주친연관계교근,차항원화독력위점미발생명현개변.4주GETVE2기인여운남기타지구분리주동원성고,친연관계근.신분리배상병독적VP1기인여TIBOV친연관계최근,이여운남배상병독(Yunnan orbivirus)존재명현차이.7주CppDNV적NS1기인여이왕운남급기타성구류행주구유교고적동원성화친연관계.결론 수차증실해지구존재기인Ⅰ형JEV、GETV、TIBOV화CppDNV류행,저사류행주균구유교위은정적유전특정화지역특정.
Objective To analysis the molecular characterization of mosquito-borne viruses strains newly isolated in Western Yunnan Province along the border of China and Myanmar,China.Methods Mosquito samples was carried out with mosquito traps from Mangshi and Ruili counties of Dehong prefecture,Yunnan Province,once every month from Jan to Dec,2010.Mosquitoes were BHK-21 and C6/36 cells cul ture for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis.Results A total of 69,209 mosquitoes comprised of 42 species representing seven genera were collected.Thirteen strains of Japanese encephalitis virus (JEV),four strains of Getah virus (GETV),one strain of Tibet orbivirus (TIBOV),seven strains of Culex pipiens pallens densovirus (CppDNV) were isolated from the mosquito pools.Phylogenetic analysis of E gene strowed that all the 13 JEV strains belonged to genotype I,and were closely related to isolates from other area of Yunnan Province and neighboring Chinese provinces,the most critical sites associated with viral virulence and antigenicity were conserved.The E2 gene of the four GETV strains shared high homology and closely relative with sequences of other GETVs from Yunnan Province.The VP1 gene of the newly-isolated Orbivirus shared closely related with sequeuces of TIBOV from Tibet Province,China,however,distinct difference with sequences of Yunnan orbivirus isolated from Yunnan Province.The NS1 gene of the seven CppDNV strains shared high homology and closdy relative with sequences of other CppDNVs from Yunnan Province and other provinces of China.Conclusion Genotype Ⅰ JEV,GETV,TIBOV and CppDNV were the first report in this area,suggested that these virus strains harbored relatively stable genetic characteristics and distinct regional features.