作　　者： ; ; ; ; ; ;

机构地区： 中国动物疫病预防控制中心

出　　处： 《中国兽医杂志》 2008年第7期3-5,共3页

摘　　要： 根据猪流感病毒(SIV)的M基因序列设计了1对特异性引物M1/M2,扩增出大小为229 bp的目的片段。针对这个基因片段,再设计合成4条寡核苷酸探针,其中反向引物的5′端用荧光素Cy3标记。以荧光标记不对称PCR技术为基础,通过将单链PCR产物与芯片杂交实现对SIV的检测,建立SIV的基因芯片检测方法。利用该方法对39份猪组织样品进行检测,与RT-PCR检测方法相比,本方法具有良好的特异性和敏感性。试验结果表明,用该方法快速检测组织中SIV是可行的,对该病的快速诊断和分子流行病学调查具有重要意义。 According to the published sequence of swine influenza virus （SIV） M gene in GenBank, M1/ M2 primers were designed to amplify the gene （about 229 bp）. Four oligonucleotide probes were designed to capture the reverse strands of the PCR product, the specific reverse primers were labeled with fluorescein （Cy3） modification at 5＇-terminal. The Cy3-1abeled PCR products were mixed with a hybridization solution and transferred to a hybridization area of the microarray. Examination of 39 tissue samples from pigs revealed that the oligonucleotide microarray assay was more sensitive and specific than RT-PCR test. The oligonucleotide microarray was proved to be a specific, sensitive, rapid and simple method and could be used to research on epidemiology and early clinical diagnosis of SIV at a molecular level.

关 键 词： 猪流感病毒 基因芯片 检测

领　　域： [农业科学] [农业科学] [农业科学]