2013～2018年中国部分地区HH9N2亚型禽流感流行病毒HA基因序列分析
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摘要
研究旨在了解2013～2018年中国部分地区分离的69株H9N2 AIV的HA基因遗传特点和变异情况。通过对HA基因片段进行扩增、克隆、序列测定及遗传进化分析。结果显示:69株病毒的裂解位点氨基酸序列分别为PSRSSR↓GLF、PSKSSR↓GLF两种形式,均是非连续碱性氨基酸,符合低致病性AIV的特征;毒株属于h9.4.2.5分支,核苷酸相似性为83.7%～99.9%,推导的氨基酸相似性为85.8%～99.9%,与现有疫苗毒株核苷酸同源性存在较大的差异;毒株受体结合位点的7个氨基酸中第198位氨基酸的变化较大,存在V、T、K、A 4种情况;同时发现69株病毒存在6～8个不同位置的潜在糖基化位点,第218位糖基化位点的大量突变与第313位糖基化位点的增加已经成为H9N2 AIV的变异特征。提示应关注流行毒株与现有疫苗毒株的抗原匹配性和基因变化等问题。
The purpose of this paper was to understand the genetic characteristics and variation of HA gene of 69 strains of H9 N2 AIV isolated from parts of China from 2013 to 2018. The HA gene fragment was amplified, cloned,sequenced and genetically evolved. The results showed that the amino acid sequences of the cleavage sites of 69 strains were PSRSSR↓GLF and PSKSSR↓GLF, both of which were non-continuous basic amino acids, which wereconsistent with the characteristics of low pathogenic H9 N2 AIV; the strain belonged to h9.4.2.5 branches, nucleotidehomology was 83.7% to 99.9%, deduced the similarity of amino acid was 85.8% to 99.9%, there was a big differencein the similarity of nucleotide with existing vaccine strains; The 198 th amino acid changed greatly. At the same time,69 strains of potential glycosylation sites in 6 to 8 different positions were found. A large number of mutations atthe 218 th glycosylation site and an increase in the 313 th glycosylation site had become characteristic of the variation ofH9 N2 AIV. It was suggested that attention should be paid to the antigen matching and genetic changes of the prevalentstrains and the existing vaccine strains.
The purpose of this paper was to understand the genetic characteristics and variation of HA gene of 69 strains of H9 N2 AIV isolated from parts of China from 2013 to 2018. The HA gene fragment was amplified, cloned,sequenced and genetically evolved. The results showed that the amino acid sequences of the cleavage sites of 69 strains were PSRSSR↓GLF and PSKSSR↓GLF, both of which were non-continuous basic amino acids, which wereconsistent with the characteristics of low pathogenic H9 N2 AIV; the strain belonged to h9.4.2.5 branches, nucleotidehomology was 83.7% to 99.9%, deduced the similarity of amino acid was 85.8% to 99.9%, there was a big differencein the similarity of nucleotide with existing vaccine strains; The 198 th amino acid changed greatly. At the same time,69 strains of potential glycosylation sites in 6 to 8 different positions were found. A large number of mutations atthe 218 th glycosylation site and an increase in the 313 th glycosylation site had become characteristic of the variation ofH9 N2 AIV. It was suggested that attention should be paid to the antigen matching and genetic changes of the prevalentstrains and the existing vaccine strains.
引文
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[2]郭元吉,谢健屏,王敏,等.从我国人群中再次分离到H9N2亚型流感病毒[J].中华实验和临床病毒学杂志,2000,14(3):209-212.
[3]GUO Y J,XIE J P,WANG M,et al.A strain of influenza A H9N2 virus repeatedly isolated from human population in China[J].Chinese J Exp Clin Virol,2000,14(3):209-212.
[4]GU M,CHEN H,LI Q,et al.Enzootic genotype S of H9N2avian influenza viruses donates internal genes to emerging zoonotic inf1uenza viruses in China[J].Vet Microbiol,2014,174(3/4):309-315.
[5]刘金华,吴清民,陈福勇,等.鸡源株与人源株H9N2流感病毒血凝素受体结合位点氨基酸比较与分析[J].中国预防兽医学报,2003,25(6):416-418.
[6]黄欣梅,李银,刘宇卓,等.华东地区H9N2亚型禽流感病毒HA基因的遗传演化分析[J].江苏农业学报,2015,31(2):382-388.
[7]JIANG W,LIU S,HOU G,et al.Chinese and global distribution of H9 subtype avian influenza viruses[J].PLoSOne,2012,7(12):e52671.
[8]PU J,WANG S,YIN Y,et al.Evo1ution of the H9N2 influenza genotype that facilitated the genesis of the novel H7N9 virus[J].Proc Natl Acad Sci,2015,112(2):548-553.
[9]LAM T T,WANG J,SHEN Y,et al.The genesis and source of the H7N9 inf1uenza viruses causing human infections in China[J].Nature,2013,502(7470):241-244.
[10]LIU J,ZHANG J,HUANG F,et al.Complex reassortment of po1ymerase genes in Asian influenza A virus H7and H9 subtypes[J].Infect Genet Evol,2014,23(2):203-208.