科研動態(tài)

為驗證自行研制的A型流感病毒分型基因芯片方法在實際中的應(yīng)用效果，了解A型流感病毒各亞型在我國不同地區(qū)和不同宿主中的分布情況，對我國20個地區(qū)、不同宿主源（雞、鴨、鵪鶉、鴿子、豬、人等）拭子和組織樣品共16 852份進行了檢測。利用自行研制的A型流感病毒分型基因芯片方法，對樣品進行RNA提取、多重不對稱RT-PCR擴增、芯片雜交、洗滌和掃描分析；同時將基因芯片檢測陽性的樣品，用普通RT-PCR擴增后進行序列測定。結(jié)果顯示：利用該方法檢出陽性樣品2 582份，總檢出率為15.32%（2 582/16 852），共檢測到H1N1、H2N9、H3N2、H4N6、H5N1、H7N1、H7N9、H9N2、H10N5、H16N3等10個亞型，且均與陽性樣品測序結(jié)果一致。結(jié)果表明，該方法可準確檢測不同地區(qū)、不同宿主中的A型流感病毒不同亞型。同時，該方法能在同一張芯片上準確鑒定A型流感病毒的多種亞型，解決了其他常規(guī)方法無法檢測已發(fā)現(xiàn)和可能發(fā)生變異的所有亞型的棘手問題，從而為A型流感診斷和分子流行病學監(jiān)測提供了一種新技術(shù)。

Application of Influenza A Virus Genotyping Microarray in Surveillance of Influenza Virus Subtypes

In order to verify the effect of influenza A virus genotyping microarray in practice，and to make clear the distribution of all subtypes of influenza A virus in various hosts in different regions in China，16 852 swabs and tissue samples collected from different hosts（chickens，ducks，quails，dove，pigs and human）in 20 regions were tested，and then RNA extraction，multiple asymmetric RT-PCR amplification，chip hybridization，cleaning and scanning analysis were carried out by the self-developed genotyping microarray. Meanwhile，the positive samples were amplified by conventional RT-PCR followed by sequencing. The results showed that 2 582 positive samples were detected out，with a total detection rate of 15.23%（2 582/16 852），involving 10 subtypes including H1N1，H2N9，H3N2，H4N6，H5N1，H7N1，H7N9，H9N2，H10N5 and H16N3，which was consistent with the result of conventional RT-PCR. It was concluded that different subtypes of influenza A virus could be detected in various hosts in different regions in China by the genotyping microarray that could also accurately identify various subtypes in the same microarray and solve the difficulty in detecting all identified or variant subtypes compared to the conventional methods，so a new technology was developed for confirmation of influenza A virus and molecular epidemiological surveillance.

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