H5亞型和N6亞型禽流感病毒二重RT-PCR檢測方法的建立
發(fā)布時間:2018-02-25 01:07
本文關(guān)鍵詞: 禽流感病毒 H亞型 N亞型 二重逆轉(zhuǎn)錄-聚合酶鏈反應(yīng) 出處:《動物醫(yī)學(xué)進展》2017年03期 論文類型:期刊論文
【摘要】:為了建立一種快速、簡便的H5亞型、N6亞型禽流感病毒(AIV)的檢測方法,根據(jù)GenBank中H5亞型、N6亞型AIV的HA和NA基因保守序列,分別設(shè)計了2對特異性引物,通過優(yōu)化條件,建立了H5亞型和N6亞型AIV二重RT-PCR檢測方法。特異性試驗結(jié)果顯示,該方法對H5N6亞型AIV特異性擴增出418bp和251bp目的片段,對H5Ny(y≠6)亞型AIV擴增出418bp目的片段,對HxN6(x≠5)亞型AIV擴增出251bp目的片段,對其他亞型和常見的禽病病原體均未擴增出目的片段;敏感性結(jié)果顯示,該方法對H5亞型和N6亞型最低檢測限為1.59×10-5ng/μL。本研究建立的H5亞型和N6亞型AIV檢測方法,具有特異性強,靈敏度高的特點,為H5亞型和N6亞型AIV臨床檢測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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