基因Ⅰ/Ⅴ型日本脑炎嵌合病毒的构建及其生物学特性和致病性研究  

Construction of Gene I/V Japanese Encephalitis Chimeric Virus and Its Biological Characteristics and Pathogenicity Research

作　　者：夏琦琦 张俊杰 杨扬 张妍 赵秋华 李宗杰 李蓓蓓[1] 刘珂[1] 邵东华[1] 邱亚峰[1] 魏建超[1] 马志永[1] XIA Qiqi;ZHANG Junjie;YANG Yang;ZHANG Yan;ZHAO Qiuhua;LI Zongjie;LI Beibei;LIU Ke;SHAO Donghua;QIU Yafeng;WEI Jianchao;MA Zhiyong(Shanghai Veterinary Research Institute,CAAS,Shanghai 100141,China;Yangtze University,Jingzhou 434000,China;Minhang Center of Animal Disease Prevention and Control in Shanghai,Shanghai 201109,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海2002412 [2]长江大学,荆州434000 [3]上海市闵行区动物疫病预防控制中心,上海201109

出　　处：《中国动物传染病学报》2023年第2期44-50,共7页Chinese Journal of Animal Infectious Diseases

基　　金：中央级公益性科研院所基本科研业务费专项(Y2020PT40,2019JB10);上海市科委农业领域项目(17391901600)。

摘　　要：为了构建基因Ⅰ/Ⅴ型日本脑炎嵌合病毒并对其生物学特性和致病性进行研究,本研究通过反向遗传技术,以GI型JEV SD12-F120株为骨架获得了含有GV型JEV XZ0934株结构蛋白的嵌合病毒,进一步通过间接免疫荧光、生长曲线、蚀斑和动物试验对其生物学特征和致病性进行了鉴定。结果显示,嵌合病毒拯救成功,与亲本病毒相比,其在BHK-21细胞上的复制效率具有显著差异,而蚀斑大小显著大于亲本病毒(P<0.001)。采用脑内和腹腔注射小鼠测得嵌合病毒的半数致死量(LD_(50))分别为10^(-2.2)PFU和10^(-1.5)PFU,表明该嵌合病毒具有强神经毒力和神经侵袭力,为后续GV型JEV的致病机制和疫苗的研究奠定了基础。In order to construct a gene type I/V Japanese encephalitis chimeric virus and analyze its biological characteristics and pathogenicity,a chimeric virus containing the structural protein of the GV JEV XZ0934 strain was obtained by reverse genetic technology using the GI JEV SD12-F120 strain as the backbone.The biological characteristics and pathogenicity of this chimeric virus were analyzed using indirect immunofl uorescence,growth curve,plaque and animal experiments.The results showed that the successful rescue of the Chimeric virus was identified by the indirect immunofl uorescence test.As compared with the parent virus,its replication efficiency on BHK-21 cells was significantly different,and the plaque size was significantly larger than the parent virus(P<0.001).The Chimeric virus was injected into mice via intracerebral and intraperitoneal routes and the LD_(50)was measured to be 10-2.2 PFU and 10^(-1.5)PFU,indicating its strong neurovirulence and neuroinvasiveness.These results laid a solid foundation for future research on vaccine development and pathogenic mechanism of JEV.

关 键 词：日本脑炎病毒 基因型 反向遗传技术 嵌合病毒 半数致死量 

分 类 号：S852.65[农业科学—基础兽医学]