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鲤春病毒血症病毒新型液相芯片检测技术的建立
引用本文:尹伟力,方绍庆,刘宁,岳志芹,刘荭,耿金培,许红岩.鲤春病毒血症病毒新型液相芯片检测技术的建立[J].兽医大学学报,2013(12):1813-1817.
作者姓名:尹伟力  方绍庆  刘宁  岳志芹  刘荭  耿金培  许红岩
作者单位:[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]深圳出入境检验检疫局,广东深fiI518000
基金项目:基金项目:山东检验检疫局科研目计划(SK201005);国家质检总局科技计划资助项目(20121K018);公益性行业科研专项经费资助项目(201210055-4)
摘    要:为建立检测鲤春病毒血症病毒(SVCV)的液相芯片快速检测技术,用DNAStar软件对GenBank中SVCV的G基因进行序列分析,设计SVCV特异性引物和探针并标记生物素,偶联荧光编码微球后与SVCV病毒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的检测出SVCV。病毒核酸的最低检出量为10pg,检测特异性高,初步建立了检测SVCV的液相芯片技术,为进一步构建其他水生动物病原体全新快速高通量检测平台奠定基础。

关 键 词:鲤春病毒血症病毒  液相芯片  检测

Development of a new liquid chip detection technique for detecting spring viraemia of carp virus
Authors:YIN Wei-li  FANG Shao-qing  LIU Ning  YUE Zhi-qin  LIU Hong  GENG Jin-pei  XU Hong-yan
Institution:1. Yantai Entry-exit Inspection and Quarantine Bureau, Yantai , Shandong 264000, China Shandong Entry-exit Inspection and Quarantine Bureau, Qingdao , Shandong 266000, China Shenzhen Entry-exit Inspection and Quarantine Bureau, Shenzhen, Guangdong 518000, China)
Abstract:To develop liquidchip technique for detecting spring viraemia of carp virus(SVCV). The G gene of SVCV in the GenBank was sequenced by using the software DNAStar 7. 0. Specific SVCV probe labeled with biotin was prepared and coupled with fluorescence-coded microspheres. The probe was used for hybridization reaction to PCR products of SVCV, then liquidchip detection technique for detection of SVCV was established by using BD FACSArray to detect fluorescence signal in the reaction system. The results showed that SVCV could be accurately detected by liq- uidchip detection technique. The lowest detection limit with this method was 10 pg of viral RNA, and the specificity of this method was very high. The liquidchip detection technique to detect SVCV was established preliminarily, which provided a foundation for further research on rapidly high throughput detection for other aquatic pathogens with this technique.
Keywords:spring viraemia of carp virus  liquichip  detection
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