2019年陕西省三带喙库蚊中淡色库蚊浓核病毒的分离与鉴定  

Isolation and identification of Culex pipiens pallens densovirus from Culex tritaeniorhynchus in Shaanxi province in 2019

作　　者：徐学平 付士红[2] 李彬 应思超 聂凯[2] 李兴洲[1] 王环宇[2] 梁国栋[2] Xu Xueping;Fu Shihong;Li Bin;Ying Sichao;Nie Kai;Li Xingzhou;Wang Huanyu;Liang Guodong(School of Public Health,Jiamusi University,Jiamusi 154007,China;Department of Arboviruses,National Institute for Viral Disease Control and Prevention,State Key Laboratory for Infectious Disease Prevention and Control,Chinese Center for Disease Control and Prevention,Beijing 100052,China)

机构地区：[1]佳木斯大学公共卫生学院,佳木斯154007 [2]中国疾病预防控制中心病毒病预防控制所虫媒室传染病预防控制国家重点实验室,北京100052

出　　处：《中华实验和临床病毒学杂志》2022年第2期225-229,共5页Chinese Journal of Experimental and Clinical Virology

基　　金：传染病预防控制国家重点实验室重点项目(2015SKLID505)。

摘　　要：目的本研究对2019年陕西省三带喙库蚊标本中分离的病毒(SX1943)进行病毒学和分子生物学鉴定。方法蚊虫研磨液接种组织培养细胞,对分离物进行病毒学和病毒分子遗传学分析。结果在2019年于陕西省采集的三带喙库蚊标本中获得1株病毒分离物(SX1943),将该病毒接种至C6/36细胞后第3天发生明显细胞病变,表现为细胞圆缩、聚集和脱落等,分离物可稳定传代。然而,该批标本在BHK-21细胞连续传代3次均未发现显著细胞病变。SX1943病毒接种至C6/36细胞后经电子显微镜(负染)观察,可见大量圆形病毒颗粒,直径约25 nm。SX1943病毒基因组扩增与测序结果显示,该病毒基因组序列全长为3594 nt,共编码3种蛋白,分别为非结构蛋白(NS1和NS2)和衣壳蛋白(VP),3种蛋白的基因长度分别为2376 nt、1098 nt和1136 nt。经病毒分子遗传进化分析发现,SX1943病毒与浓核病毒亚科Brevidensovirus病毒属中的淡色库蚊浓核病毒(Culex pipiens pallens Densovirus,CppDNV)处于同一进化分支,上述结果提示SX1943病毒为CppDNV。结论陕西省三带喙库蚊中分离到CppDNV。Objective To perform virological and molecular biological identification of the virus(SX1943)isolated from specimens of Culex tritaeniorhynchus in Shaanxi province in 2019.Methods Mosquito ground fluid was inoculated into tissue culture cells,and isolates were made virological and molecular genetic analysis.Results A virus isolate(SX1943)was obtained from a specimen of Culex tritaeniorhynchus collected in Shaanxi province in 2019,which developed significant cytopathic effect(CPE)on day 3 after inoculation into C6/36 cells,manifested as cell pyknosis,aggregation and shedding,and the isolate could be stably passaged.However,no significant CPE was observed after three consecutive passages in BHK-21 cells.Electron microscopy(negative staining)of C6/36 cells inoculated with SX1943 virus showed a large number of round virus particles,about 25 nm in diameter.The result of amplification and sequencing of the SX1943 viral genome showed that the total length of the viral genome sequence was 3594 nt,encoding a total of three proteins,which were non-structural proteins(NS1 and NS2)and capsid proteins(VP),and the gene lengths of the three proteins were 2376 nt,1098 nt and 1136 nt,respectively.Phylogenetic analysis of the virus revealed that the SX1943 virus were in the same evolutionary clade as Culex pipiens pallens Densovirus(CppDNV)in the genus Brevidensovirus in the subfamily Densovirus,and the above result suggested that SX1943 virus was CppDNV.Conclusions CppDNV was isolated from Culex tritaeniorhynchus in Shaanxi province.

关 键 词：淡色库蚊浓核病毒 三带喙库蚊 系统进化分析 

分 类 号：R373.3[医药卫生—病原生物学]