H10亚型和N8亚型禽流感病毒双重RT-PCR检测方法的建立

发布时间：2018-04-19 12:27

  本文选题：禽流感病毒 + H亚型　； 参考：《畜牧与兽医》2017年01期

【摘要】：根据Gen Bank中H10亚型和N8亚型禽流感病毒(AIV)的HA和NA基因保守序列,分别设计筛选出2对特异性引物,用于H10亚型和N8亚型AIV的检测,优化引物之间的浓度,建立了同时检测H10亚型和N8亚型AIV的双重RT-PCR检测方法。该方法对H10N8亚型AIV可特异性扩增出267 bp(H10亚型)和464 bp(N8亚型)目的条带,对H10Ny(y≠8)亚型AIV仅扩增出267 bp目的条带,对HXN8(x≠10)亚型AIV仅扩增出464 bp目的条带,对其他亚型AIV和常见禽病病原体均未扩增出任何条带。该方法对H10亚型和N8亚型AIV检测下限均为104拷贝数/μL。本研究建立的H10亚型和N8亚型AIV双重RT-PCR特异性强、灵敏度高,可同时快速检测H10亚型和N8亚型AIV,为其感染的快速鉴别诊断提供一种简便、快速和有效的方法。
[Abstract]:According to the conserved sequences of HA and na genes of H10 subtype and N8 subtype avian influenza virus in Gen Bank, two pairs of specific primers were designed and screened for the detection of H10 subtype and N8 subtype AIV, and the concentration between primers was optimized.A dual RT-PCR method for simultaneous detection of H 10 subtype and N 8 subtype AIV was established.The target bands of H10N8 subtype AIV and 464 bp(N8 subtype were amplified by this method, only 267bp for H10Ny(y 鈮，

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