摘要
目的对辽宁省朝阳市蚊虫中分离的一种新病毒进行分子生物学鉴定。方法在辽宁省朝阳市采集蚊虫,用细胞培养方法分离病毒,对引起C6/C36细胞病变(CPE)的病毒进行逆转录-聚合酶链式反应(RT-PCR)检测,并对阳性PCR产物进行测序分析;对病毒编码区全基因进行测序并与GenBank中36株黄病毒参考毒株序列进行比较;用Mega3.1软件、以邻位相连法构建黄病毒属编码区全基因遗传进化树。结果从蚊虫中分离到2株病毒,黄病毒属通用引物RT-PCR检测均为阳性,扩增片段大小986bp,其中有600～750bp核苷酸碱基序列与登革病毒2型、伊桂普病毒、塞皮克病毒和班奇病毒有66%～68%的一致性,2株病毒间碱基序列100%一致,为同一种黄病毒;病毒编码区基因序列全长10308bp,与其他黄病毒比较结果表明,分离株E基因、NS3基因及NS5基因碱基序列与其他黄病毒碱基序列一致性最高分别为59.6%,61.7%和67.0%;编码区全基因遗传进化树分析结果显示,分离株与登革病毒群、日本脑炎病毒群等蚊传黄病毒拥有共同进化祖先,但在系统发生树上处在单独进化分支。结论辽宁省朝阳市分离到的黄病毒毒株不属于现有黄病毒属任何一种群,为黄病毒属的一个新种群,经GenBank分析确认为世界首次发现,并命名为朝阳病毒,收录号为FJ883471。
Objective To identify a new virus of flavivirus by molecular biology, which was isolated from mosquitoes collected in Chaoyang of Liaoning province. Methods Mosquitoes were collected in Chaoyang of Liaoning province and the virus was isolated by cell culture. The isolated virus causing cytopathogenic effect（CPE） on C6/C36 cells was tested by reverse transcription polymerase chain reaction（RT-PCR） and the positive PCR-product was sequenced and analyzed. Coding sequence（CDS） compIete genome of the virus was sequenced and compared with the sequences of 36 isolates from Gen- Bank. Phylogenetic tree of CDS complete genome of flavivirus were constructed by neighbor-joining method in Mega 3.1 soft ware. Results Two viral swains were isolated from the mosquitoes and the results of RT-PCR were positive by using universal primers of flavivirus. The length of amplified segment was 986 bp,600 -750 bp of which was similar to Dengue virus 2,Iguape virus,Sepik virus and Banzi virus and the nucleotide identities ranged between 66% and 68%. The nucleotide identifies between two isolates were 100% and so they actually were one flavivirus. The length of the nucleotide sequence of CDS complete genome of the virus was 10 308 bp and identifies of E gene,NS3 gene and NS5 gene were 59. 6% ,61.7% and 67.0% same,respectively,with the other flavivirus in GenBank. Phylogenetic analysis of CDS complete genome of flavivirus showed the isolate together with Dengue virus group, Japanese encephalitis virus group, Kokobera virus group, Ntaya virus group and Spondweni virus group belonged to one ancestor, but the isolate had its own branch in the tree. Conclusion The isolate from mosquitoes in Chaoyang of Liaoning province belongs to flavivirus, but it is not a member of any other group of flavivirus and it constructs a new group of flavivirus. The new virus is first reported in the world and named Chaoyang virus. The GenBank accession number is FJ883471.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2009年第7期769-772,共4页
Chinese Journal of Public Health
基金
辽宁省博士科研基金项目(20052135)
关键词
黄病毒
朝阳病毒
新种群
flavivirus
Chaoyang virus
new species
