病毒性出血性败血症病毒液相芯片检测技术的建立

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Gen-Bank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.