云南环状病毒(Yunnan orbivirus,YOUV)在牛与蚊上的分离鉴定  

Isolation and Identification of Yunnan OrbivirusS(YOUV)from Cattle and Mosquitoes

作　　者：杨恒 王怡丹 董双平 姚萍芬 付余飞 周明 王宵 王斌 牛保生 陈伟 YANG Heng;WANG Yidan;DONG Shuangping;YAO Pingfen;FU Yufei;ZHOU Ming;WANG Xiao;WANG Bin;NIU Baosheng;CHEN Wei(School of Agriculture and Life Sciences,Kunming University,Kunming 650214,China;College of Animal Science and Technology,College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;Animal Disease Control Center of Shizong County,Qujing 655700,China;Agricultural and Rural Technology Comprehensive Service Center of Wu Long Village,Qujing 655700,China;Medical School,Kunming University of Science and Technology,Kunming 650500,China)

机构地区：[1]昆明学院农学与生命科学学院,昆明650214 [2]华中农业大学动物科技学院动物医学院,武汉430070 [3]云南省师宗县动物疫病预防控制中心,曲靖655700 [4]五龙乡农业农村技术综合服务中心,曲靖655700 [5]昆明理工大学医学院,昆明650500

出　　处：《病毒学报》2023年第3期752-763,共12页Chinese Journal of Virology

基　　金：昆明学院引进人才科研项目(项目号:YJL2210),题目:蓝舌病毒衣壳蛋白VP2基因修饰型溶瘤病毒抗胃癌调节作用机制研究。

摘　　要：云南环状病毒(Yunnan orbivirus, YUOV)为环状病毒属新成员,病毒于1997年首次从我国云南省采集的三带喙库蚊(Culex tritaeniorhynchus)中分离。随后YOUV相继在秘鲁的发病马(1997)、日本的哨兵牛(2018)以及美国病死的白尾鹿(2019)中分离,提示该病毒分布广泛,可能是一种危害动物健康的虫媒病毒。2022年我们在云南省师宗县采集的牛血液与蚊样本中分离出三株在白纹伊蚊细胞(C6/36)上引起明显细胞病变的病毒。电镜下观察,病毒粒子具有环状病毒属病毒的形态特征;琼脂糖凝胶电泳结果显示,病毒基因组由多节段双链RNA组成。对环状病毒属高度保守的基因节段及其编码蛋白(Seg-2/T2)序列分析显示,分离毒株在系统发生树上与YOUV聚为一簇,序列相似度在82.7%/98.1%~96.7%/99.8%(nt/aa)之间,表明分离三株病毒为YOUV成员;对决定环状病毒属血清型的基因节段及其编码蛋白(Seg-3/OC1)序列分析显示,分离毒株与已知YOUV毒株的序列相似度在84.5%/97.4%~90.3%/95.7%之间,提示目前世界范围发现YOUV均属同一血清型。为分析YOUV在当地牛羊中的流行情况,采用本研究建立的YOUV qRT-PCR方法对45份采集至师宗县牛羊血液进行检测,结果显示病毒在羊上的感染率为5%(1/20),在牛上的感染率达24%(6/25);感染动物血清中YOUV的中和抗体效价在1:16至1:215之间。本研究首次在云南省的蚊与牛血液中同时分离出YOUV,证实了病毒在师宗县牛羊中的流行。我国流行YOUV的遗传特征、感染动物宿主范围与致病性有待进一步研究。Yunnan orbivirus(YOUV),a new member of genus Orbivirus,was first isolated from mosquito(Culer tritaeniorhynchus)collected from Yunnan Province,China in 1997.Later,the virus was isolated from sick horse in Peru(1997),sentinel cattle in Japan(2018)and white-tailed-deer died of disease in USA(2019),indicating that YOUV might be an arbovirus threaten animal health.In this study,three strains of viruses inducing strong cytopathic effect on inoculated aedesalbopictus(C6/36)cells were isolated from cattle blood and mosquito samples collected in Shizong County of Yunnan Province in 2022,China.Electron microscope observation revealed that the virus particles have morphological feature of orbiviruses,while the viral genome was composed of multi segments dsRNA as revealed through agarose gel electrophoresis.Sequence analysis of highly conserved protein and its encoding segment(Seg-2/T2)of orbiviruses indicated that the isolated viruses clustered together with other reported YOUV strains in the phylogenetic tree with sequence identities of nucleotide and amino acid(nt/a)ranging from 82.7%/98.1%to 96.7%/99.8%.Sequence analysis of protein to determining the serotype and its encoding segment(OC1/Seg3)of orbiviruses showed that the isolated viruses exhibited sequence identities ranging from 84.5%/97.4%to 90.3%/95.7%with other YOUV strains,indicating that only one serotype of YOUV was found around the world.In order to investigated the prevalence of YOUV in local cattle and goats,YOUV specific qRT-PCR and RT-PCR methods were developed in this study.A total of 45 blood samples collected from cattle and goats in Shizong County were screened for the infection of YOUV with the prevalence of 5%(1/20)for goats and 24%(6/25)for cattle.The exposures of the herds to YOUV were further confirmed by serum neutralization test using blood samples tested positive by qRT-PCR,with antibody titers ranging from 1:16 to 1:215.In this study,YOUV was first isolated from mosquitoes and the blood of cattle in Yunnan Province,China,confirming the prevale

关 键 词：环状病毒属 云南环状病毒 序列分析 流行病学调查 

分 类 号：S852.65[农业科学—基础兽医学]