黄鑫儿, 周雪, 侯赛, 侯银续, 俞俊岭, 陈晴晴, 祝茜, 何兰, 龚磊, 吴家兵, 何军. 2020年安徽省1株候鸟源H5N8禽流感病毒遗传进化特征分析[J]. 疾病监测, 2022, 37(1): 115-122. DOI: 10.3784/jbjc.202106030319
引用本文: 黄鑫儿, 周雪, 侯赛, 侯银续, 俞俊岭, 陈晴晴, 祝茜, 何兰, 龚磊, 吴家兵, 何军. 2020年安徽省1株候鸟源H5N8禽流感病毒遗传进化特征分析[J]. 疾病监测, 2022, 37(1): 115-122. DOI: 10.3784/jbjc.202106030319
Huang Xin′er, Zhou Xue, Hou Sai, Hou Yinxu, Yu Junling, Chen Qingqing, Zhu Qian, He Lan, Gong Lei, Wu Jiabing, He Jun. Genetic and evolutionary characteristics of an avian influenza A (H5N8) virus isolated from migratory bird[J]. Disease Surveillance, 2022, 37(1): 115-122. DOI: 10.3784/jbjc.202106030319
Citation: Huang Xin′er, Zhou Xue, Hou Sai, Hou Yinxu, Yu Junling, Chen Qingqing, Zhu Qian, He Lan, Gong Lei, Wu Jiabing, He Jun. Genetic and evolutionary characteristics of an avian influenza A (H5N8) virus isolated from migratory bird[J]. Disease Surveillance, 2022, 37(1): 115-122. DOI: 10.3784/jbjc.202106030319

2020年安徽省1株候鸟源H5N8禽流感病毒遗传进化特征分析

Genetic and evolutionary characteristics of an avian influenza A (H5N8) virus isolated from migratory bird

  • 摘要:
      目的  分析安徽省肥东县候鸟监测站检测的H5N8亚型禽流感病毒(AIVs)的遗传进化特征。
      方法  从收集的死亡天鹅样本中提取病毒RNA,使用实时荧光定量RT-PCR区分流感亚型,并采用一代测序方法对阳性样本的HA和NA基因扩增后测序,应用生物信息学软件构建系统发育进化树并分析其分子特征。
      结果  2020年12月1日收到来自安徽省肥东县1只死亡天鹅的鼻拭子、鸟喙拭子和泄殖腔拭子样本,PCR结果均显示H5阳性,测序结果提示3份样本为同一株H5N8亚型禽流感病毒。 遗传进化分析显示,该病毒属于2.3.4.4支系且与埃及、俄罗斯等地分离株有很高的相似性,其HA与NA基因都在欧亚谱系中,并且已经适应了不同宿主。 该病毒HA裂解位点呈现高致病性分子特征,存在6个糖基化位点,且发生S133L、S137A、T160A、D187N、K193N、Q196K、S227R氨基酸突变。 PB2蛋白发生K389R、V589T突变、 PA蛋白发生N409S突变、 M1蛋白发生N30D、T215A突变、NS1蛋白发生P42S突变。
      结论  本次发现的H5N8亚型禽流感病毒受体结合位点仍为禽源受体,但具有人源受体分子特征,其他氨基酸位点都提示该病毒有毒力增强的危险性,需进一步加强监测。

     

    Abstract:
      Objective  To analyze the genetic and evolutionary characteristics of an avian influenza A (H5N8)virus strain isolated from a dead swan at a migratory bird monitoring station in Feidong county of Anhui province.
      Methods  Virus RNA extraction was conducted by using the samples collected from the dead swan, and real-time fluorescence quantitative RT-PCR was used the subtyping of influenza virus. HA and NA gene sequencing after amplification of the positive samples were conducted by first-generation sequencing method. Bioinformatics software was used to construct phylogenetic evolutionary tree and analyze its molecular characteristics.
      Results  The nasal swab, beak swab and cloacal swab from the dead swan in Feidong, which were received on December 1, 2020, were all positive for H5 subtype by PCR. Sequencing results showed that the strains from three samples shared the same sequence of avian influenza A (H5N8) virus. Phylogenetic analysis showed that the virus belonged to 2.3.4.4 lineage and had high similarity with the isolates from Egypt and Russia. The HA and NA genes were in Eurasian lineage and had been adapted to different hosts. The HA cleavage site of the virus showed highly pathogenic molecular characteristics, carried 6 glycosylation sites and contained the amino acid mutations of S133L, S137A, T160A, D187N, K193N, Q196K and S227R. There were K389R and V589T mutations in PB2 protein, N409S mutations in PA protein, N30D and T215A mutations in M1 protein, S31N mutations in M2 protein, and P42S mutations in NS1 protein.
      Conclusion  The receptor binding site of avian influenza A (H5N8) virus isolated in this study was still avian origin receptor, but it had the molecular characteristics of human origin receptor. Mutations in other amino acid sites suggested the risk of increased virulence of avian influenza virus, and further surveillance is needed.

     

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