摘要
根据GenBank发表的H9亚型禽流感病毒(AIV(H9))HA基因、新城疫病毒(NDV)F基因和传染性支气管炎病毒(IBV)N基因分别设计了3对特异性引物,通过优化扩增条件,建立了同时检测AIV(H9)、NDV和IBV的多重PCR方法,对其特异性和敏感性进行检验,并在临床中进行了初步应用。结果表明,该多重PCR方法具有良好的特异性和敏感性,可同时扩增出大小分别为380bp(AIV(H9))、529bp(NDV)和853bp(IBV)的特异片段,能分别检出1pg AIV(H9)、1pg NDV和10pg IBV的RNA模板,且对AIV(H9)、NDV和IBV混合感染的临床阳性病料的检测结果与单项PCR的检测结果一致。该多重PCR方法可为AIV(H9)、NDV和IBV的临床检测和流行病学调查提供技术支撑。
Three pairs of specific primers were designed according to the HA gene ot H9 avian mHuenza vi- rus （AIV（Hg））, F gene of Newcastle disease virus （NDV） and N gene of infectious bronchitis virus （IBV） in GenBank, and the multiplex PCR for simultaneous detection of AIV（H9）, NDV and IBV was es- tablished by optimizing the reaction conditions. The specificity and sensitivity were tested and some clinical samples were tested by the multiplex PCR. The specific gene fragments of 380 bp （AIV（H9））, 529 bp （NDV） and 853 bp （IBV） were amplified. The specificity and sensitivity of the multiplex PCR were good, with as little as 1 pg of AIV （H9） and NDV, and 10 pg of IBV could be detected in RNA templates. The results detected by the multiplex PCR were consistent with the results detected by single PCR for the mixed positive samples containing AIV（H9）, NDV and IBV. The developed multiplex PCR can provide the important method for diagnosis and epidemiological investigation in clinic.
出处
《动物医学进展》
CSCD
北大核心
2014年第1期12-16,共5页
Progress In Veterinary Medicine
基金
青岛市公共领域科技支撑计划(10-3-3-17-nsh)
作者简介
徐守振（1977-），男，山东沂水人，副教授，博士，主要从事动物禽病防治研究。
