摘要
利用多重反转录聚合酶链式反应同时检测H5、H7和H9亚型禽流感病毒.在GenBank中搜索H5、H7和H9亚型禽流感病毒(AIV)的血凝素基因序列,并利用DNAStar软件分析其相似性,利用Primer Premier 5.0软件设计3对分别针对AIV H5、H7和H9亚型的特异性引物.这3对引物所扩增的cDNA片段大小分别为427、228和830 bp.结果表明,通过对多重RT-PCR扩增条件的优化,建立了同时检测H5、H7和H9亚型AIV的多重RT-PCR技术.该多重RT-PCR对H5、H7和H9亚型AIV能同时扩增出3条大小分别为427、228和830 bp的cDNA片段,与其他常见禽病病原的核酸不存在交叉反应.该多重RT-PCR对H5、H7和H9亚型AIV cDNA的最低检出量分别为10 pg、1 ng和10 pg.
Hemagglutinin gene sequences of H5, H7 and H9 subtypes avian influenza virus were searched in GenBankTM, and their similarity were analyzed using DNAStar. Three sets of specific oligonucleotide primers were designed. Specific cDNA bands of 427,228 and 830 bp were observed simultaneously for RNA isolated from HS, H7 and H9 subtypes AIV. There was no cross-reaction with other avian pathogens after amplification. The minimal amount of cDNA as detected by this multiplex RT-PCR was 10 pg cDNA of H5,1 ng cDNA of H7 and 10 pg cDNA of H9 subtypes AIV.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2009年第3期95-98,共4页
Journal of South China Agricultural University
基金
吉林省科技发展计划项目(20065020)
吉林农业大学青年启动基金项目资助
