2014～2016年江苏地区H9N2亚型禽流感病毒遗传进化及其分子特征的分析
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摘要
H9N2亚型禽流感病毒(avian influenza virus, AIV)在我国广泛流行,给养禽业造成了巨大的经济损失,还对公共卫生安全存在潜在威胁. 2014～2016年, H9N2亚型禽流感病毒在江苏地区的活禽市场的鸡中的阳性率为8.39%～13.74%.选取10株H9N2分离株进行全基因组测序及遗传进化分析.结果表明, H9N2分离株的HA基因与早期的江苏分离株位于同一小分支上,且同源性较高,但NA基因与早期江苏病毒株的同源性较低,核苷酸同源性为91.4%～93.9%,氨基酸同源性为91.2%～93.8%.这些病毒株的基因型依然为B69,为目前的优势基因型.所有H9N2病毒分离株的HA蛋白的226位点为L.固相受体结合实验结果显示, H9N2分离株具有人源α2,6唾液酸受体的结合特性.
H9N2 avian influenza virus(AIV) circulates widely in China, causing a large economic loss to poultry industry and posing a big potential threat to public health. From 2014 to 2016, the positive ratio of H9N2 AIV isolation from chicken in the living market of Jiangsu province is 8.39%–13.74%. Ten H9N2 virus isolates were selected for whole genome sequencing and phylogenetic analysis.The result demonstrated that the HAs of those H9N2 virus isolates belonged to the same lineage and had a high homology with earlier Jiangsu isolates. In contrast, NA genes of those isolates evolved far away from the earlier Jiangsu isolates. Compared to earlier isolates, the nucleotide homology of NA genes and amino acid homology of NA protein were ranged from 91.4% to 93.9% and 91.2% to 93.8%, respectively. All those H9N2 viruses belonged to genotype B69 which predominates at present in China. The result of MegAlign analysis showed that 226 site in the hemagglutinins of those H9N2 isolates were Leucine. Further, it was proved that those H9N2 virus isolates preferred to bind to human type α2,6 sialic acid receptor in solid-phase receptor binding assay.
H9N2 avian influenza virus(AIV) circulates widely in China, causing a large economic loss to poultry industry and posing a big potential threat to public health. From 2014 to 2016, the positive ratio of H9N2 AIV isolation from chicken in the living market of Jiangsu province is 8.39%–13.74%. Ten H9N2 virus isolates were selected for whole genome sequencing and phylogenetic analysis.The result demonstrated that the HAs of those H9N2 virus isolates belonged to the same lineage and had a high homology with earlier Jiangsu isolates. In contrast, NA genes of those isolates evolved far away from the earlier Jiangsu isolates. Compared to earlier isolates, the nucleotide homology of NA genes and amino acid homology of NA protein were ranged from 91.4% to 93.9% and 91.2% to 93.8%, respectively. All those H9N2 viruses belonged to genotype B69 which predominates at present in China. The result of MegAlign analysis showed that 226 site in the hemagglutinins of those H9N2 isolates were Leucine. Further, it was proved that those H9N2 virus isolates preferred to bind to human type α2,6 sialic acid receptor in solid-phase receptor binding assay.
引文
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14 Horimoto T,Kawaoka Y.Pandemic threat posed by avian influenza A viruses.Clin Microbiol Rev,2001,14:129-149
15 Wan H,Perez D R.Amino acid 226 in the hemagglutinin of H9N2 influenza viruses determines cell tropism and replication in human airway epithelial cells.J Virol,2007,81:5181-5191
16 Li C,Yu K,Tian G,et al.Evolution of H9N2 influenza viruses from domestic poultry in Mainland China.Virology,2005,340:70-83
17 Aamir U B,Wernery U,Ilyushina N,et al.Characterization of avian H9N2 influenza viruses from United Arab Emirates 2000 to 2003.Virology,2007,361:45-55
18周锦萍,李凯航,鞠厚斌,等.上海地区活禽批发市场H9亚型禽流感病毒调查.中国动物传染病学报,2010,18:56-61
19 Shinya K,Ebina M,Yamada S,et al.influenza virus receptors in the human airway.Nature,2006,440:435-436
20 Kobasa D,Kodihalli S,Luo M,et al.Amino acid residues contributing to the substrate specificity of the influenza A virus neuraminidase.J Virol,1999,73:6743-6751
21 Pu J,Sun H,Qu Y,et al.M gene reassortment in H9N2 influenza virus promotes early infection and replication:contribution to rising virus prevalence in chickens in China.J Virol,2017,91:e02055
2陈伯伦,张泽纪,陈伟斌.禽流感研究:Ⅰ鸡A型流感病毒分离与血清学初步鉴定.中国兽医杂志,1994,20:3-5
3吴海燕,李明义,范根成,等.33株H9N2亚型禽流感病毒分离株分子流行特别的研究.中国动物检疫,2010,27:31-33
4 Li X,Shi J,Guo J,et al.Genetics,receptor binding property,and transmissibility in mammals of naturally isolated H9N2 Avian Influenza viruses.PLoS Pathog,2014,10:e1004508
5 Pu J,Wang S,Yin Y,et al.Evolution of the H9N2 influenza genotype that facilitated the genesis of the novel H7N9 virus.Proc Natl Acad Sci USA,2015,112:548-553
6 Peiris M,Yuen K,Leung C,et al.Human infection with influenza H9N2.Lancet,1999,354:916-917
7 Butt K M,Smith G J D,Chen H,et al.Human infection with an avian H9N2 influenza A virus in Hong Kong in 2003.J Clin Microbiol,2003,43:5760-5767
8 Xu J,Li S,Yang Y,et al.Human infection with a further evolved avian H9N2 influenza A virus in Sichuan,China.Sci China Life Sci,2018,61:604-606
9 Wang J,Wu M,Hong W,et al.Infectivity and transmissibility of avian H9N2 influenza viruses in pigs.J Virol,2016,90:3506-3514
10 Chen H Y,Yuan H,Gao R,et al.Clinical and epidemiological characteristics of a fatal case of avian influenza A H10N8 virus infection:a descriptive study.Lancet,2014,383:714-721
11 Gao R,Cao B,Hu Y,et al.Human infection with a novel avian-origin influenza A(H7N9)virus.N Engl J Med,2013,368:1888-1897
12 Dong G,Luo J,Zhang H,et al.Phylogenetic diversity and genotypical complexity of H9N2 influenza A viruses revealed by genomic sequence analysis.PLoS ONE,2011,6:e17212
13 Teng Q,Xu D,Shen W,et al.A single mutation at position 190 in hemagglutinin enhances binding affinity for human type sialic acid receptor and replication of H9N2 avian influenza virus in mice.J Virol,2016,90:9806-9825
14 Horimoto T,Kawaoka Y.Pandemic threat posed by avian influenza A viruses.Clin Microbiol Rev,2001,14:129-149
15 Wan H,Perez D R.Amino acid 226 in the hemagglutinin of H9N2 influenza viruses determines cell tropism and replication in human airway epithelial cells.J Virol,2007,81:5181-5191
16 Li C,Yu K,Tian G,et al.Evolution of H9N2 influenza viruses from domestic poultry in Mainland China.Virology,2005,340:70-83
17 Aamir U B,Wernery U,Ilyushina N,et al.Characterization of avian H9N2 influenza viruses from United Arab Emirates 2000 to 2003.Virology,2007,361:45-55
18周锦萍,李凯航,鞠厚斌,等.上海地区活禽批发市场H9亚型禽流感病毒调查.中国动物传染病学报,2010,18:56-61
19 Shinya K,Ebina M,Yamada S,et al.influenza virus receptors in the human airway.Nature,2006,440:435-436
20 Kobasa D,Kodihalli S,Luo M,et al.Amino acid residues contributing to the substrate specificity of the influenza A virus neuraminidase.J Virol,1999,73:6743-6751
21 Pu J,Sun H,Qu Y,et al.M gene reassortment in H9N2 influenza virus promotes early infection and replication:contribution to rising virus prevalence in chickens in China.J Virol,2017,91:e02055