关键词: 禽流感病毒, 基因芯片, 亚型鉴定

Abstract: A rapid microarray-based assay for the reliable detection of H5, H7 and H9 subtypes of avian influenza virus (AIV) was developed. The strains used in the experiment were A/Turkey/England/N28/73（H5N2），A/African starling/983/79（H7N1）and A/Turkey/Wiscosin/1/66（H9N2）. The cDNAs clone which encode approximately 500bp avian influenza virus gene fragments were obtained by reverse transcription PCR technique, the DNA fragments, which were reamplified from recombinant plasmids, were spotted on a glass-bound microarray. Cy5-labeled fluorescent probes were hybridized to these samples cDNAs, which generated from vRNA during reverse transcription. These samples cDNAs contained multiple fragments of AIV including the hemagglutinin gene and matrix gene. The type of AIV were detected with the matrix gene，and the subtypes of AIV were differentiated with hemagglutinin gene. The arrays were scanned to determine the probe binding sites. The hybridization pattern agreed approximately with the known grid location of each target. The results showed that DNA microarray technology could provide a useful diagnostic method for AIV.

Key words: Avian influenza virus, Microarray, Subtype identification