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Molecular and phylogenetic analysis of matrix gene of avian influenza viruses isolated from wild birds and live bird markets in the USA

机译:从美国野生鸟类和活禽市场分离的禽流感病毒基质基因的分子和系统发育分析

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Background Wild birds are the natural hosts for influenza A viruses (IAVs) and provide a niche for the maintenance of this virus. Objectives This study was undertaken to analyze nucleotide sequences of the matrix (M) gene of AIVs isolated from wild birds and live bird markets (LBMs) to index the changes occurring in this gene. Methods M-gene of 229 avian influenza virus (AIV) isolates obtained from wild birds and LBMs was amplified and sequenced. Full-length sequences (~900nt.) thus obtained were analyzed to identify changes that may be associated with resistance to adamantanes. Phylogenetic analysis of all sequences was performed using clustalw, and evolutionary distances were calculated by maximum composite likelihood method using mega (ver. 5.0) software. Results Twenty-seven different viral subtypes were represented with H3N8 being the most dominant subtype in wild birds and H7N2 being the predominant subtype among isolates from LBMs. Phylogenetic analysis of the M-gene showed a high degree of nucleotide sequence identity with US isolates of AIVs but not with those of Asian or European lineages. While none of the isolates from wild birds had any antiviral resistance-associated mutations, 17 LBM isolates carried polymorphisms known to cause reduced susceptibility to antiviral drugs (adamantanes). Of these 17 isolates, 16 had S31N change and one isolate had V27A mutation. Conclusions These results indicate independent evolution of M-gene in the absence of any antiviral drugs leading to mutations causing resistance indicating the need for continued active surveillance of AIVs.
机译:背景野生鸟类是A型流感病毒(IAV)的天然宿主,并为维持这种病毒提供了利基。目的进行本研究以分析从野生鸟类和活禽市场(LBM)分离的AIV的基质(M)基因的核苷酸序列,以索引该基因中发生的变化。方法对野生鸟类和LBMs获得的229株禽流感病毒(AIV)的M基因进行扩增和测序。分析由此获得的全长序列(〜900nt。)以鉴定可能与对金刚烷抗性相关的变化。使用clustalw对所有序列进行系统进化分析,并使用mega(ver。5.0)软件通过最大复合似然法计算进化距离。结果代表了27种不同的病毒亚型,其中H3N8是野生鸟类中最主要的亚型,而H7N2是来自LBM的分离株中的主要亚型。 M基因的系统发育分析表明,其与美国的AIV分离株具有高度的核苷酸序列同一性,但与亚洲或欧洲谱系却没有。尽管来自野生鸟类的分离株均未具有任何抗病毒耐药性相关突变,但17个LBM分离株具有已知导致抗病毒药物(金刚烷)敏感性降低的多态性。在这17个分离株中,有16个具有S31N改变,一个分离株具有V27A突变。结论这些结果表明在不存在任何抗病毒药物的情况下M基因的独立进化会导致引起耐药性的突变,这表明需要对AIV进行持续的主动监测。

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