病毒性出血性败血症病毒液相芯片检测技术的建立  

Development of a Liquid Chip Technique for Detection of Viral Hemorrhagic Septicemia of Salmonids Virus(VHSV)

作　　者：尹伟力 梁君妮 林超[2] 张四化 岳志芹[2] 刘荭[4] 

机构地区：[1]烟台出入境检验检疫局,山东烟台264000 [2]山东出入境检验检疫局,山东青岛266000 [3]武汉市重大动物疫病防控中心,湖北武汉430003 [4]深圳出入境检验检疫局,广东深圳518000

出　　处：《中国动物检疫》2015年第6期64-68,共5页China Animal Health Inspection

基　　金：公益性行业科研专项经费项目201210055-4;科技部十二五支撑项目2013BAD12B02;国家质检总局科技计划项目2012IK018

摘　　要：为建立检测病毒性出血性败血症病毒(VHSV)的液相芯片快速检测技术,用DNAStar软件对GenBank中VHSV的G基因进行序列分析,设计VHSV特异性引物并标记生物素。探针氨基化修饰并与荧光编码微球偶联后与VHSV RT-PCR产物杂交反应,用液相芯片仪器检测荧光信号。结果表明液相芯片检测体系能够正确检出VHSV。病毒核酸的最低检出量为10pg,检测特异性高,说明初步建立了检测VHSV的液相芯片技术,为VHSV的检测提供了新方法。In order to develop a liquid chip assay for detection of viral hemorrhagic septicemia of salmonids virus （VHSV）,the G gene of VHSV in the GenBank was sequenced by using the software DNAStar 7.0 and specific VHSV primers labeled with biotin were prepared and coupled with fluorescence-coded microspheres. The aminationed probe was used for hybridization of VHSV PCR products,and a liquid chip detection technique for detection of VHSV was established by using BD FACSArray to detect fluorescence signal in the reaction system. The results showed that VHSV could be accurately detected by the developed liquid chip detection technique. The lower detection limit with this method was 10 pg of viral RNA,and the specificity of this method was very high.The study provided a new method for rapid and high throughput detection of VHSV.

关 键 词：病毒性出血性败血症病毒(VHS) 液相芯片 检测 

分 类 号：S941.413[农业科学—水产养殖]