作　　者： ; ; ; ; ; ;

机构地区： 华南农业大学兽医学院农业部养禽与禽病防治重点开放实验室

出　　处： 《中国动物检疫》 2007年第5期25-28,共4页

摘　　要： 根据猪繁殖与呼吸综合征病毒(PRRSV)的核衣壳蛋白编码基因序列设计了一对特异性引物P1/P2,扩增出大小为294bp的目的片段;再针对这个基因片段,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PRRSV的检测,建立PRRSV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病料组织中PRRSV是可行的,对该病的进行快速诊断和分子流行病学调查具有重要意义。 According to the Gene Bank published sequence of porcine reproductive and respiratory syndrome virus （PRRSV） , P1/ P2 primes were designed to amplify the gene （about 294bp） Four oligonucleotide probes were designed to capture the reverse strand of the PCR product, the specific reverse primers were labeled with fluorescein （Cy3） modification at 5＇-terminal. The Cy3-labeled 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 in epidemiology research and early clinical diagnosis of PRRSV at molecular level.

关 键 词： 猪繁殖与呼吸综合征病毒 基因芯片

领　　域： [农业科学] [农业科学] [农业科学]