新城疫液相芯片快速检测方法的建立  

Preliminary Development of Liquichip Detection Technique for Rapid Detection of Newcastle Disease Virus

作　　者：詹爱军[1,2] 王新卫[3] 金鑫[2] 花群义[2] 卢体康[2] 陈枝楠[2] 秦爱建[1] 

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]深圳出入境检验检疫局动植物检验检疫技术中心,广东深圳518010 [3]河南农业大学牧医工程学院,河南郑州450002

出　　处：《现代生物医学进展》2009年第15期2903-2906,共4页Progress in Modern Biomedicine

基　　金："十一五"国家科技支撑项目(2006BAK20A29);深圳出入境检验检疫局科技项目(SZ2008102)资助

摘　　要：目的:建立可检测新城疫病毒(Newcastle disease virus,NDV)的液相芯片快速检测技术。方法:用DNAStar软件对GEN-BANK中NDV的NP基因进行序列分析设计NDV特异性探针并标记生物素,利用该探针与荧光编码微球偶联后与抽提的NDV病毒RNA的RT-PCR产物杂交反应,用液相芯片检测仪(Liquichip 200)检测荧光信号建立了NDV快速液相芯片检测方法。结果:检测结果显示,该法具有较好的特异性,不与H5AIV和H9AIV反应;检测灵敏度达到150个EID50;该法与鸡胚病毒分离法检出NDV的符合率达到97.1%。结论:初步建立了检测NDV的液相芯片技术,为进一步搭建NDV全新快速高通量检测平台奠定了基础,也为其他同类病毒的快速高通量检测提供了借鉴和经验。Objective： The aim is to develop Liquichip detection technique to detect Newcastle disease virus in practical poultry industry. Methods： The NP gene of NDV in the GENEBANK was sequenced by using the software DNAStar 5.0. Specific NDV probe labeled with biotin was prepared and coupled with fluorescence-coded microspheres. The probe was used for hybridization reaction to RT-PCR products of NDV, then Liquichip detection technique for detection of Newcastle disease virus was developed by using Liquichip 200 to detect fluorescence signal from the reaction system. Results： The results showed that this assay method displayed better specificity to NDV and no respond to H5 and H9 serotype avian influenza virus when being used for detection. The sensitivity test indicated the de- tecting limit for the NDV could reach to 150 EID50. The accordance rate between Liquichip detection way and virus isolation by chicken eggs was 97.1%. Conclusion： The Liquichip detection technique to detect NDV was established preliminarily, which provided a founda- tion for further research on rapidly high throughput detection for NDV and also for exploration to detect other viruses with the technique in poultry industry.

关 键 词：新城疫病毒 液相芯片 检测 

分 类 号：S855.3[农业科学—临床兽医学]