羊口疮病毒DZ株在不同细胞上的增殖特性研究  

Growth characteristics of ORFV-DZ strain in different cells

作　　者：杨倩 李鹏飞[1,2] 鲜思美 包涛涛[1,4] 张友 闵德省 梁倩 顾庆林 YANG Qian;LI Pengfei;XIAN Simei;BAO Taotao;ZHANG You;MIN Desheng;LIANG Qian;GU Qinglin(College of Animal Science,Guizhou University,Guiyang 550025,China;Animal Husbandry and Veterinary Medicine Station of Luhe District,Nanjing City,Nanjing 211500,China;Institute of Animal Disease Research of Guizhou,Guiyang 550025,China;Bureau of Agriculture and Rural Affairs of Southeast Guizhou,Kaili 556000,China)

机构地区：[1]贵州大学动物科学学院,贵阳550025 [2]南京市六合区畜牧兽医站,南京211500 [3]贵州省动物疫病研究所,贵阳550025 [4]黔东南州农业农村局,贵州凯里556000

出　　处：《扬州大学学报（农业与生命科学版）》2021年第5期19-24,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：贵州省科技计划项目(黔科合支撑[2018]2264);贵州省科学技术基金资助项目(黔科合基础[2019]1113)。

摘　　要：为研究羊口疮病毒(ORFV)DZ株在不同细胞上的增殖特性,将ORFV-DZ株分别接种犊牛睾丸细胞(BT)和鸡胚成纤维细胞(CEF)2种原代细胞,以及山羊皮肤成纤维细胞系(GSFs)、小鼠胚胎成纤维细胞系(NIH3T3)、宫颈癌细胞系(Hela)、牛肾细胞系(MDBK)、犬肾细胞系(MDCK)、非洲绿猴肾细胞系(Vero)、乳仓鼠肾细胞系(BHK-21)7种传代细胞,通过细胞病变(CPE)、间接免疫荧光试验(IFA)、TCID_(50)测定和实时荧光定量PCR检测ORFV-DZ株在不同细胞上的增殖特性。结果表明:ORFV-DZ株能在BT、Hela以及MDBK 3种细胞中有效增殖,且出现细胞变圆、聚集、脱落等CPE;IFA检测ORFV-DZ株接种这3种细胞均能出现特异性绿色荧光;收集BT、Hela和MDBK细胞培养的F5代细胞毒,测得其TCID_(50)值分别为10^(-5.81)·0.1 mL^(-1)、10^(-4.22)·0.1 mL^(-1)、10^(-4.88)·0.1 mL^(-1);qPCR检测结果中,ORFV-DZ株在BT细胞中随传代次数增加病毒拷贝数未发生明显变化,在MDBK细胞中F5代时病毒拷贝数已下降,而在Hela细胞中,随着传代次数的增多,病毒拷贝数逐步下降。这一研究得到了适合ORFV-DZ株有效增殖的细胞为BT和MDBK,为后续ORFV的基础研究以及疫苗研发等提供了细胞学研究基础。The aim of this study was to study the proliferation characteristics of Orf virus(ORFV)DZ strain in different cells.In this study,ORFV-DZ strain was inoculated into two primary cells bovine testis cell(BT)and chicken embryo fibroblast(CEF),as well as seven cell strains or cell lines,including goat skin fibroblast cell strain(GSFs),mouse embryonic fibroblast cell strain(NIH3T3),hela cell line(Hela),mardin-darby bovine kidney cell strain(MDBK),mardin-darby canine kidney cell strain(MDCK),verda reno(Vero)and baby hamster syrian kidney cell strain(BHK-21).The proliferation characteristics of ORFV-DZ strain in different cells were detected by CPE,indirect immunofluorescence(IFA),TCID_(50) and real-time quantitative PCR.The results showed that ORFV-DZ strain could effectively proliferate in BT,HeLa and MDBK cells,and CPE such as cell roundness,aggregation and abscission appeared.IFA detects ORFV-DZ strain inoculated with these three kinds of cells can show specific green fluorescence.ORFV-DZ strain F5 generation cultures in BT,Hela and MDBK cells was collected,and the TCID_(50) values determined were 10^(-5.81),10^(-4.22),10^(-4.88) copies·0.1 mL^(-1);qPCR detection showed that ORFV-DZ proliferated in BT cells with the increase of passages,however,the virus copy number did not change significantly.In MDBK cells,the virus copy number had declined at the F5 generation,while in Hela cells,the virus copy number gradually decreased with the increase of passage generations.In this study,the cells suitable for the effective proliferation of the ORFV-DZ strain were BT and MDBK cells,which provides a cytological research basis for the follow-up basic research of ORFV and its vaccine development.

关 键 词：羊口疮病毒 细胞 增殖特性 

分 类 号：S852.65[农业科学—基础兽医学]