鸭瘟病毒在鸭胚脐带间充质干细胞上生物学特性的研究  

Biological characteristics of duck plague virus on duck embryo umbilical cord mesenchymal stem cells

作　　者：徐丽晶[1,2] 陆涛峰[2] 王怡平 陈洪岩[2] 赵丽丽[2] 

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]中国农业科学院,哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江省实验动物与比较医学重点实验室,黑龙江哈尔滨150069

出　　处：《中国兽医科学》2017年第9期1094-1099,共6页Chinese Veterinary Science

基　　金：国家科技支撑计划项目(2015BAI07B02-02);中国农业科学院基本科研业务费专项(Y2016PT41);黑龙江省自然科学基金重点项目(ZD2016006);中央级公益性科研院所基本科研业务费专项(1610302016013,1610302017013)

摘　　要：为了解鸭瘟病毒(DPV)在鸭胚脐带间充质干细胞上的适应性和增殖情况。本试验将DPV接种鸭胚脐带间充质干细胞,连续传代至第10代,通过CPE观察、病毒粒子电镜观察、核酸检测、间接免疫荧光检测等分析其生物学特性,并测定了连续10代的病毒滴度(TCID_(50))。结果显示,第一代DPV即在鸭胚脐带间充质干细胞上产生了明显的CPE,并且在连续传代的过程中CPE能够稳定出现。初步提纯的病毒悬浮液在电镜下可见到结构清晰的子代病毒。F1~F10代DPV均能扩增出大小为416 bp的UL6基因片段。间接免疫荧光又进一步证实DPV的阳性血清能够对染毒病灶产生特异性荧光染色。连续测定了10代的病毒滴度(TCID_(50))均不低于10^(-6.43),且在DEUCMSC上测定的DPV TCID_(50)明显高于DEF的。说明DPV能在鸭胚脐带间充质干细胞上稳定增殖。In order to understand the adapt ability and proliferation of duck plague virus （DPV） in duck embryonic umbilical cord mesenchymal stem cells,DPV virus＇ s biological characteristics in the duck embryo umbilical cord mesenchymal stem cells after the continuous passage for 10 generations was analyzed by observation of CPE and electron microscopy,nucleic acid detection,indirect immunofluorescence detection,and determination of TCID（50） of the continuous passage of 10 generations.Lesions were seen by the results of CPE duck plague virus in the duck embryo umbilical cord mesenchymal stem cells on the first generation,and they were stable even after several generations.The progeny virus could be seen under electron microscopy.The fragment of UL6 gene was amplified with a size of 416 bp.It was further confirmed that the positive serum of against DPV could be specific fluorescent staining for infected lesions in indirect immunofluorescence analysis.The virus titer of the continuous 10 generations was not less than 10^-6.43.The titer（TCID50） which was determined by DPV on DEUCMSC was significantly higher than that of DEF.It was proved that DPV could proliferate on duck embryonic umbilical cord mesenchymal stem cells.

关 键 词：鸭瘟病毒 鸭胚脐带间充质干细胞 生物学特性 

分 类 号：S852.659.1[农业科学—基础兽医学]