H5亚型和N6亚型禽流感病毒二重RT-PCR检测方法的建立

发布时间：2018-02-25 01:07

  本文关键词： 禽流感病毒 H亚型 N亚型 二重逆转录-聚合酶链反应　出处：《动物医学进展》2017年03期 　论文类型：期刊论文

【摘要】：为了建立一种快速、简便的H5亚型、N6亚型禽流感病毒(AIV)的检测方法,根据GenBank中H5亚型、N6亚型AIV的HA和NA基因保守序列,分别设计了2对特异性引物,通过优化条件,建立了H5亚型和N6亚型AIV二重RT-PCR检测方法。特异性试验结果显示,该方法对H5N6亚型AIV特异性扩增出418bp和251bp目的片段,对H5Ny(y≠6)亚型AIV扩增出418bp目的片段,对HxN6(x≠5)亚型AIV扩增出251bp目的片段,对其他亚型和常见的禽病病原体均未扩增出目的片段;敏感性结果显示,该方法对H5亚型和N6亚型最低检测限为1.59×10-5ng/μL。本研究建立的H5亚型和N6亚型AIV检测方法,具有特异性强,灵敏度高的特点,为H5亚型和N6亚型AIV临床检测以及防控提供了有效方法。
[Abstract]:In order to establish a rapid and simple method for the detection of H5 subtype N6 avian influenza virus, two pairs of specific primers were designed according to the conserved sequences of HA and na genes of H5 subtype N6 subtype AIV in GenBank. A double RT-PCR method for the detection of H5 subtype and N6 subtype AIV was established. The results of specific test showed that 418 BP and 251 BP target fragments were amplified by this method for H 5N 6 subtype AIV, and 418 BP target fragment was amplified for H 5N 6 subtype AIV. For HxN6(x 鈮，

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