禽流感病毒液相芯片检测方法的建立

Development of a suspension array method for the detection of avian influenza virus

针对禽流感病毒M基因设计了特异性引物和探针,将不对称RT-PCR扩增产物和偶联到磁性微球上的探针进行杂交,通过检测荧光值,建立了检测禽流感病毒通用亚型的液相芯片方法,并对该方法进行了特异性、敏感性、稳定性试验以及田间样品检测试验。结果表明,该方法能特异地检出禽流感病毒,病毒核酸的最低检出量为10-4 ng,与新城疫病毒、禽传染性支气管炎病毒等常见病毒无交叉反应。该方法具有检测准确,特异性强、灵敏度高、简便快速的优点,为一种快速、准确、高通量检测动物疫病的新方法。

The aim of the present study was to develop a suspension array to detect avian influenza virus（AIV）. The gene M of AIV deposited in the GenBank was analyzed to design primers which were lablled by biotin during synthesis. A pair of specific probes labelled with amine was prepared and coupled with fluorescencecoded microspheres. The probes were used for hybridization reaction to RT-PCR product. The liquid detection system-based suspension array was established with the specificity, sensitivity and stability validated by using AIV strains and samples from different areas. The results showed that AIV could be identified with high sensitivity,which could reach up to 10-4 ng. And no cross reaction was found with other common avian viruses, such as Newcastle disease virus, and infectious bronchitis virus. The result showed that the suspension array technique established in the present study had good specificity,sensitivity and accuracy, which provided foundation and exploration for further research in the detection of other viruses.