爲驗證自行研制的A型流感病毒分型基因小店芯片方法在實際中的應用效果，了解A銀城型流感病毒各亞型在我國(guó)不同地區和不同化日宿主中的分布情況，對(duì)我國(guó)20道男個地區、不同宿主源（雞、鴨、鹌鹑、鴿子、豬、人等）拭子和組織樣喝鐵(yàng)品共16 852份進(jìn)行了檢測。利用自行研制的物商A型流感病毒分型基因芯片方法，對(duì)樣(yàng少唱)品進(jìn)行RNA提取、多哥雨重不對(duì)稱RT-PCR擴增、芯片雜交、洗滌和掃描分生區析；同時(shí)將(jiāng)讀道基因芯片檢測陽性的樣(yàng)品，用普子弟通RT-PCR擴增後(hòu)進(jìn)行序列測定。結果坐木顯示：利用該方法檢出陽性樣(yà為慢ng)品2 582份，總檢出率爲1見黑5.32%（2 582/16 852），共檢測到H1N1、H2N9、都音H3N2、H4N6、H5N1、H對冷7N1、H7N9、H9N2、H10N5、H16N3等10個亞型，且均與陽性樣(飛女yàng)品測序結果一緻。結果表明，該方法可準确檢子我測不同地區、不同宿主中的A型流感病毒不同亞型。同紙日時(shí)，該方法能(néng)在同一張芯片上準确鑒定A型流感病毒們好的多種(zhǒng)亞型，解決了其他常規方法無法檢測已務電發(fā)現和可能(néng)發(fā)生變異的所有亞訊弟型的棘手問題，從而爲A型流感診斷和分子流行病學(xué)監測在河提供了一種(zhǒng)新技術。
Application of Influenza A Virus北東 Genotyping Microarray in化現 Surveillance of Influ唱北enza Virus Subtypes
In order to verify the effec短醫t of influenza 器對A virus genotyping microarray in practi劇西ce，and to make clear the distribution 員聽of all subtypes of influenza A vi歌裡rus in various hosts in d店外ifferent regions in China，16 852問年 swabs and tissue samples collect都新ed from different腦媽 hosts（chickens，ducks，quails，慢要dove，pigs and human）in 20 regions 年哥were tested，and鄉家 then RNA extract短雜ion，multiple asymmetric低爸 RT-PCR amplification，chip hybridiza工機tion，cleaning and scannin物站g analysis were carried out by the s土視elf-developed g微湖enotyping micro工暗array. Meanwhile，the positive samples 錯場were amplified by conv如員entional RT-PCR 校就followed by sequencing. The花離 results showed that 2 582 positive s數子amples were detecte事身d out，with a total detecti大長on rate of 15.23%（2 582/16 計問852），involving 10 subtypes好文 including H1N1，H2N9，H3N2，H4N6，H5N好小1，H7N1，H7N9，H9N2做請，H10N5 and H16N3，which was consi厭從stent with the res制能ult of conventional RT-PCR. It was co購冷ncluded that dif吃們ferent subtypes of influenz場唱a A virus could be dete房大cted in various hosts i林低n different regions in China by the ge年物notyping microarray that could a放街lso accurately identify various sub聽朋types in the same microar去化ray and solve the 下新difficulty in det照說ecting all identified or variant subt冷一ypes compared to the conven錢海tional methods，so a new technolog北高y was developed for co你窗nfirmation of influenz很拍a A virus and molecular epid海機emiological surveillance.
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