鲤春病毒血症病毒新型液相芯片检测技术的建立

Development of a new liquid chip detection technique for detecting spring viraemia of carp virus

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。

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.