一株进口胎牛血清源BVDV毒株的分离与鉴定  

Isolation and identification of a BVDV strain from imported fetal bovine serum

作　　者：温树波 宋扬 蒙小刚 陈卓 孟令浩 王沐笙 王云龙 霍晓伟[1] 刘锴[1] 翟景波[2,3] 鲁会军 金宁一 WEN Shu-bo;SONG Yang;MENG Xiao-gang;CHEN Zhuo;MENG Lin-hao;WANG Mu-sheng;WANG Yun-long;HUO Xiao-wei;LIU Kai;ZHAI Jin-bo;LU Hui-jun;JIN Ning-yi(College of Animal Science and Technology,Inner Mongolia University for Nationalities,Tongliao,Inner Mongolia 028000,China;Institute of Innovative Research on Zoonoses,Inner Mongolia University for Nationalities;Brucellosis Prevention and Treatment Technology Research Center,Inner Mongolia Autonomous Region;Military Veterinary Institute,The Academy of Military Medical Science;Khorchin Inner Mongolia Beef Cattle Breeding Co.Ltd)

机构地区：[1]内蒙古民族大学动物科技学院,内蒙古通辽028000 [2]内蒙古民族大学人兽共患病创新研究院 [3]内蒙古自治区布鲁氏菌病防治工程技术研究中心 [4]军事科学院军事医学研究院军事兽医研究所 [5]内蒙古科尔沁肉牛种业股份有限公司

出　　处：《中国病原生物学杂志》2021年第7期743-746,共4页Journal of Pathogen Biology

基　　金：内蒙古自治区布鲁氏菌病防治工程技术研究中心开放课题(No.MDK2019082);国家重点研发计划项目(No.2018YFD0500803);内蒙古自治区科技重大专项(No.2019ZD006);内蒙古民族大学博士基金启动项目(No.BS584)。

摘　　要：目的分离进口胎牛血清中的牛病毒性腹泻-粘膜病病毒(BVDV)毒株并进行系统进化分析。方法通过RT-PCR检测购买胎牛血清中的BVDV核酸,确定有BVDV污染后。将该血清样品接种MDBK细胞,连续传代5次,利用免疫荧光、PCR扩增、测序以及遗传进化分析等方法对其进行鉴定。结果成功分离到1株BVDV毒株,PCR和免疫荧光检测均为BVDV阳性,命名为FBS-TL20。基于5’UTR、Npro和E2基因序列的同源性分析表明,FBS-TL20毒株与分离自埃及新生羊体内的毒株BSU1同源性分别为99.18%、98.89%和97.27%。遗传进化分析显示,该毒株属于BVDV-1b型。结论从进口胎牛血清中成功分离到1株非致细胞病变型BVDV 1 b亚型毒株,为进一步研究BVDV的相关分子机制提供了实验材料。Objective To isolate, identify, and perform phylogenetic tree analysis of a bovine viral diarrhea-mucosal disease virus(BVDV) strain detected in imported fetal bovine serum. Methods RT-PCR was used to detect nucleic acids of BVDV in purchased fetal bovine serum, and it was therefore confirmed to be contaminated with BVDV. The serum sample was inoculated into MDBK cells and no cytopathic effect was observed after 5 passages. The isolate was identified using immunofluorescence, amplification with PCR, and genetic evolutionary analysis. Results A strain of BVDV, designated FBS-TL20, was successfully isolated and fetal bovine serum tested positive for the strain according to PCR and immunofluorescence. The TCID50 of the virus was 10-6/ml. Sequence homology analysis based on the 5’UTR, Npro, and E2 sequences indicated that the FBS-TL20 strain was most similar to the BSU1 strain(99.18% for the 5’UTR sequence, 98.89% for Npro, and 97.27% for E2), which was isolated from newborn sheep in Egypt. Genetic evolutionary analysis suggested that the strain belonged to BVDV-1 b. Conclusion A non-cytopenic strain of the BVDV 1 b subtype was successfully isolated from imported fetal bovine serum. This serum could serve as a material with which to study molecular mechanisms related to BVDV in future research.

关 键 词：牛病毒性腹泻病毒 分离鉴定 遗传进化分析 

分 类 号：S852.65[农业科学—基础兽医学]