%0 Journal Article
%T Microarray for detecting and subtyping of circulating influenza A virus<br>甲型流感病毒流行毒株检测和分型基因芯片的研制
%A Xihan Li~
%A Yu Wang~
%A Fangzheng Liu~
%A Hongwei Gu~
%A Yonghua Yang~
%A <br>李稀罕
%A 王宇
%A 刘方政
%A 顾宏伟
%A 杨永华
%J 微生物学报
%D 2010
%I 
%X Abstract: Objective] We designed and characterized an oligonucleotide microarray for detecting and subtyping of circulating influenza A virus including subtypes H1N1, H1N2, H3N2, H5N1 and H9N2. Methods] Based on the sequences of influenza A viruses obtained from the Influenza Virus Resource database of National Center for Biotechnology Information, 46 oligonucleotides probes and one quality control probe were designed to fabricate the microarray. The full-length cDNAs of hemagglutinin and neuraminidase genes were amplified by RT-PCR using universal primers, and the resulting PCR products were labeled and fragmented using Klenow fragment before hybridized with the microarray. A total of 18 different influenza A virus strains representing 5 subtypes and 186 clinical samples were used to validate the specificity and sensitivity of the microarray. Results] All 18 strains were accurately detected and subtyped by the microarray and no cross hybridization could be detected. The limit of detection for the microarray was approximately 1×104 gene copies of in vitro transcribed RNA. Of the 186 clinical samples, 8 were successfully subtyped as H1N1 and 4 were subtyped as H3N2. Conclusion] The results show that the microarray is a useful diagnostic method with high specificity and sensitivity, and could be used for influenza surveillance.
%K Influenza A virus
%K microarray
%K diagnostic<br>关键词：甲型流感病毒
%K 基因芯片
%K 检测
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=8247DE71881F41CFA07B1A14ED845316&yid=140ECF96957D60B2&vid=771152D1ADC1C0EB&iid=38B194292C032A66&sid=8ACD9060100C26F1&eid=EB58C3052341AAA3&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=0