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我国新分离芒市病毒基因组特征与细胞感染特性研究OA北大核心CSTPCD

Genomic and cellular infection characteristics of a newly isolated Mangshi virus in China

中文摘要英文摘要

目的 获取我国新发现芒市病毒(Mangshi virus,MSV)基因组特征以及在细胞上的感染特性,为开展MSV进化与致病性方面研究奠定基础.方法 通过高通量测序技术获取MSV毒株V301/YNJH/2019全基因组序列,使用IQtree、RPD4与Simplot等软件进行系统发育树的构建与基因重配分析;测定病毒在白伊蚊细胞(C6/36)、非洲绿猴肾细胞(Vero)与仓鼠肾细胞细(BHK)上的增殖特性,通过血清中和试验调查当地牛羊上病毒的感染情况.结果 病毒基因组全长20 623 bp,由12节段的双链RNA(Seg-1至Seg-12)组成.序列分析显示V301/YNJH/2019为基因重配毒株,病毒的Seg-1与Seg-11与我国湖水沉积物中发现的MSV对应基因节段序列高度相似,而Seg-2至Seg-10以及Seg-12则与我国蚊上分离的MSV毒株具有最近的亲缘关系.病毒在C6/36与BHK细胞上可高效增殖并引起细胞明显的细胞病变,在Vero细胞上可低水平复制但不引起细胞病变.在芒市采集的20份羊血清样本中未检测到MSV的中和抗体,但在采集20份牛血清中检测到2份阳性样本,其中和抗体效价分别为1:128与1:54.结论 V301/YNJH/2019为基因重配毒株,病毒可感染C6/36、BHK与Vero细胞,在芒市牛群中检测到感染动物.

The genomic characteristics and cellular tropism of a Mangshi virus(MSV)isolated in China were investigated,thereby establishing a robust foundation for further research on the evolution and pathogenicity of MSV.The genome sequence of MSV strain V301/YNJH/2019 was obtained by next-generation sequencing,followed by phylogenetic tree construction and rearrangement assessment using software IQtree,RPD4,and Simplot.Viral proliferation was assessed in C6/36(Aedes albop-ictus),Vero(African green monkey kidney),and BHK(baby hamster kidney)cells.An initial epidemiological investigation of MSV in local cattle and goats was conducted using the serum neutralization test.The genome of MSV strain V301/YNJ H/2019 was 20623 bp in length,encompassing 12 segments of double-stranded RNA(Seg-1 to Seg-12).Sequence analysis confirmed genomic rearrangement of the Seg-1 and Seg-11 sequences,ex-hibiting high similarity to MSV isolated from lake sediment in China in 2022,while Seg-2 to Seg-10 and Seg-12 were most closely related to a MSV strain isolated from mosquitoes in China in 2013.The virus efficiently proliferated and induced sig-nificant cytopathic effects(CPE)in both C6/36 and BHK cells,but limited replication and no observable CPE in Vero cells.No detectable neutralizing antibodies against MSV were detected in 20 goat serum samples collected in Mangshi,while 2 of 20 bo-vine serum samples were positive with neutralizing antibody titers of 1:128 and 1:54.Whole genome sequencing revealed re-assortment events of the V301/YNJH/2019 strain,which is capable of infecting C6/36,BHK,and Vero cells.MSV infection was confirmed in cattle in Mangshi.

杨恒;李占鸿;肖雷;李卓然;谢佳蕊;廖德芳;高林;李华春

昆明学院,农学与生命科学学院,昆明 650214||云南省畜牧兽医科学院,云南省热带亚热带动物病毒病重点实验室,昆明 650224云南省畜牧兽医科学院,云南省热带亚热带动物病毒病重点实验室,昆明 650224

畜牧业

芒市病毒;全基因组测序;基因重配;细胞感染特性;流行病学调查

Mangshi virus;complete genome sequencing;reassortment;celltropism;epidemiological survey

《中国人兽共患病学报》 2024 (006)

504-511,528 / 9

国家自然科学基金(No.31760744);国家公益性行业(农业)专项(No.201303035);昆明学院引进人才科研项目(No.YJL2210) Supported by National Natural Science Foundation of China(No.31760744);Special Fund for Agro-scientific Research in the Public Interest(No.201303035);Scientific research project for talents introduced by Kunming University(No.YJL2210)

10.3969/j.issn.1002-2694.2024.00.087

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