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当前位置: 首页 > 文章 > 蓝舌病毒14型的分离与鉴定 中国兽医科学 2019 (7) 855-860
Position: Home > Articles > 蓝舌病毒14型的分离与鉴定 Chinese Veterinary Science 2019 (7) 855-860

蓝舌病毒14型的分离与鉴定

作  者：
曹雨;张莹辉;赵炜;陈新;薄宗义;姚文生;JUNG Yong-Sam;戴建君;钱莺娟;印春生
单  位：
南京农业大学动物医学院教育部动物健康与食品安全国际合作联合实验室农业部细菌学重点实验室
关键词：
蓝舌病毒;14型;分离;鉴定
摘  要：
为了解我国新疆地区羊蓝舌病的流行情况,对某羊场疑似发生蓝舌病的羊病料进行了采集,并进行病毒分离和鉴定.应用间接ELISA试剂盒检测血清抗体,抗凝血样品接种敏感的BHK21细胞进行连续盲传,对细胞病变呈阳性的样品进行间接免疫荧光鉴定,同时针对VP7片段和VP2片段设计引物进行RT-PCR检测鉴定,并对其L2基因进行系统发育树分析.结果显示,血清中BTV抗体检测均为阳性,接种BHK21细胞后均产生了特异性细胞病变,并与FITC标记的抗BTV单抗特异性结合,针对VP7扩增出1 156 bp片段,针对VP2分别扩增出850 bp和1 581 bp片段,经测序和序列分析确定为BTV-14型.结果表明,我国新疆地区存在蓝舌病毒14型,这也是我国境内首次分离到蓝舌病毒14型,为进一步防控我国蓝舌病疫情提供了科学依据.
作  者：
CAO Yu;ZHANG Ying-hui;ZHAO Wei;CHEN Xin;BO Zong-yi;YAO Wen-sheng;JUNG Yong-Sam;DAI Jian-jun;QIAN Ying-juan;YIN Chun-sheng;MOE Joint International Research Laboratory of Animal Health and Food Safety/Key Laboratory of Animal Bacteriology,Ministry of Agriculture/College of Veterinary Medicine,Nanjing Agricultural University;China Institute of Veterinary Drug Control;
关键词：
bluetongue virus;;serotype 14;;isolation;;identification
摘  要：
In order to understand the prevalence of sheep bluetongue disease in Xinjiang,suspected bluetongue samples on a sheep farm were collected and subjected to virus isolation and identification.To this end,several experiments were performed,including indirect ELISA to determine BTV antibodies in sera,isolation of BTV by continuous blind passage on sensitive BHK21 monolayer cells which were inoculated with blood with anticoagulant and subsequently indirect immunofluorescence to examine the cytopathic positive samples,RT-PCR was used to detect and identify BTV VP7 and VP2 genes with respective primers,and phylogenetic tree analysis of L2 gene was carried out to determine the BTV serotype.In result,BTV antibodies in sera were positive,the suspicious samples inoculated on BHK21 cells produced specific cytopathic effects by blind passage and were specifically recognized by FITC labeled anti-BTV monoclonal antibody.A 1 156 bp fragment was amplified for VP7,and 850 bp and 1 581 bp fragments were amplified for VP2,respectively,and identified as BTV-14 by sequencing and sequence analysis.We concluded that this is the first time to isolate a serotype 14 bluetongue virus in Xinjiang,China,which provide scientific basis for further prevention and control of bluetongue disease in China.

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