用液相芯片方法检测禽流感病毒H5N1亚型的研究

目的基于液相芯片（MASA）技术建立一种对H5N1亚型禽流感病毒进行快速检测的新方法。方法对GenBank上已有H5和N1核酸片段进行比对分析，设计简并引物和探针，将合成的探针与荧光编码微球进行偶联，建立检测方法。利用该方法检测H5N1亚型禽流感病毒和其他常见亚型（H1N1、H2N2、H3N2、H9N2）的标本。结果利用该方法能够特异性地检测出H5N1标本，检测信号具有较高荧光强度，对其他亚型标本的检测则为阴性结果。该方法对H5NI亚型RNA的最低检出量为10pg。结论利用液相芯片技术研制的H5N1亚型禽流感病毒检测方法能够用于禽流感病毒H5N1亚型的快速、灵敏、特异性检测及鉴定。

Determination of Avian influenza virus H5N1 using MASA technology

Objective To set up a new rapid method for the rapid determination of Avian influenza virus H5N1 basing on the multi-analyte suspension array （MASA） technology. Methods Sequence analysis and design of degenerate primers and specific probes were set in the comparison and analysis of H5 and NI genes. In combination with MASA technology, these primers and probes were used for the determination of samples of HSN1 and other subtypes（ H1N1, H2N2, H3N2 and H9N2）. Results These specific primets and probes sets could be used for the rapid and specific determination of Avian influenza virus H5N1. High fluorescence intensity was detected in H5N1 sample but not in other samples of H1N1, H2N2, H3N2 and H9N2. The minimal amount as detected for HSN1 was 10 pg with this method. Conclusions MASA technology based determination method could be used for highl-specific, sensitive and rapid determination of Avian influenza virus H5N1.