导致狐狸重症腹泻疫情的新型犬冠状病毒的发现与鉴定  

Discovery and identification of a novel canine coronavirus causing a diarrhea outbreak in Vulpes

作　　者：刘雨婷 邓瑶 牛胜 朱娜 宋敬东 张晓爽 苏伟婷 佴文慧 陆柔剑 David MIrwin 高福 王文玲 王奇慧 谭文杰 张树义 Yuting Liu;Yao Deng;Sheng Niu;Na Zhu;Jingdong Song;Xiaoshuang Zhang;Weiting Su;Wenhui Nie;Roujian Lu;David M.Irwin;George Fu Gao;Wenling Wang;Qihui Wang;Wenjie Tan;Shuyi Zhang(College of Animal Science and Veterinary Medicine,Shenyang Agricultural University,Shenyang 110866,China;NHC Key Laboratory of Biosafety,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China;National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Tianjin Zoo,Tianjin 300381,China;Kunming Cell Bank,State Key Laboratory of Genetic Resources and Evolution,Kunming Institute of Zoology,Chinese Academy of Sciences,Kunming 650223,China;Department of Laboratory Medicine and Pathobiology,University of Toronto,Toronto ON M5S 1A8,Canada;CAS Key Laboratory of Pathogen Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]College of Animal Science and Veterinary Medicine,Shenyang Agricultural University,Shenyang 110866,China [2]NHC Key Laboratory of Biosafety,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China [3]College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China [4]National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China [5]Tianjin Zoo,Tianjin 300381,China [6]Kunming Cell Bank,State Key Laboratory of Genetic Resources and Evolution,Kunming Institute of Zoology,Chinese Academy of Sciences,Kunming 650223,China [7]Department of Laboratory Medicine and Pathobiology,University of Toronto,Toronto ON M5S 1A8,Canada [8]CAS Key Laboratory of Pathogen Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China [9]School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China

出　　处：《Science Bulletin》2023年第21期2598-2606,M0006,共10页科学通报（英文版）

基　　金：supported by the National Key R&D Program of China(2022YFE0210300,2022YFC2303401,2016YFD0500300,2021YFC0863300,and 2021YFC2300101);the National Natural Science Foundation of China(32070407);the special fund for Science and Technology Innovation Teams of Shanxi Province(202204051001022)。

摘　　要：2019年末,中国东北地区狐狸群中暴发大规模疫情,发病狐狸表现出急性腹泻症状,并出现大量狐狸死亡.通过对该疫源地发病狐狸肛拭子、血液与发病组织的检测分析,获得了多条全长新型犬冠状病毒(CCoV)基因组序列,研究团队将该病毒命名为VuCCoV,并确认VuCCoV是引起该疫情的病原.进一步分析表明,VuCCoV基因组的4个结构基因(S,E,M,N)中的3个(E,M,N)与犬冠状病毒(CCoV)对应基因具有超过90%的核苷酸同一性,但S基因核苷酸序列表现出较大的差异.VuCCoV的RNA依赖的RNA聚合酶与蝙蝠冠状病毒PB025的对应蛋白仅存在2~3个氨基酸差异,系统进化分析提示VuCCoV很可能来源于蝙蝠.表面等离子体共振分析实验表明,VuCCoV刺突蛋白受体结合区(RBD)可与犬和狐狸的氨肽酶N(APN)结合,但不与人APN结合,假病毒感染及病毒分离培养实验进一步证实该VuCCoV不易感染人源细胞.蝙蝠等野生动物宿主的病毒跨物种传播对人类和家畜健康构成了显著威胁,VuCCoV的发现和溯源证实蝙蝠作为疫情出现的潜在驱动因素的重要性.积极监控蝙蝠及其他犬科动物中的病毒感染,发现、鉴定对人兽健康构成潜在威胁的新型冠状病毒并研究其进化变异特征与感染特性,对于防控新发人兽共患传染病、建设国家生物安q全体系具有重要意义.Cross-species transmission of viruses from wildlife animal reservoirs,such as bats,poses a threat to human and domestic animal health.Previous studies have shown that domestic animals have important roles as intermediate hosts,enabling the transmission of genetically diverse coronaviruses from natural hosts to humans.Here,we report the identification and characterization of a novel canine coronavirus(VuCCoV),which caused an epidemic of acute diarrhea in Vulpes(foxes)in Shenyang,China.The epidemic started on November 8,2019,and caused more than 39,600 deaths by January 1,2022.Full-length viral genomic sequences were obtained from 15 foxes with diarrhea at the early stage of this outbreak.The VuCCoV genome shared more than 90%nucleotide identity with canine coronavirus(CCoV)for three of the four structural genes,with the S gene showing a larger amount of divergence.In addition,67%(10/15)of the VuCCoV genomes contained an open reading frame(ORF3)gene,which was previously only detected in CCoV-I genomes.Notably,VuCCoV had only two to three amino acid differences at the partial RNA-dependent RNA polymerase(RdRp)level to bat CoV,suggesting a close genetic relationship.Therefore,these novel VucCoV genomes represent a previously unsampled lineage of cCoVs.We also show that the VuCCoV spike protein binds to canine and fox aminopeptidase N(APN),which may allow this protein to serve as an entry receptor.In addition,cell lines were identified that are sensitive to VucCoV using a pseudovirus system.These data highlight the importance of identifying the diversity and distribution of coronaviruses in domestic animals,which could mitigate future outbreaks that could threaten livestock,public health,and economic growth.

关 键 词：犬冠状病毒 人兽共患传染病 病毒感染 表面等离子体共振 犬科动物 动物宿主 氨肽酶N 狐狸 

分 类 号：S852.65[农业科学—基础兽医学]