2009-2017年河北省廊坊市新甲型H1N1流感病毒基因进化分析
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摘要
目的了解2009-2017年河北省廊坊市新甲型H1N1流感流行态势,分析H1N1血凝素(hemagglutinin,HA)和神经氨酰酶(neuraminidase,NA)基因变异情况及其遗传进化特征。方法选定2009-2017年河北省廊坊市分离的甲型H1N1流感毒14株,使经核酸提取和PCR扩增测序HA和NA基因,采用MEGA version 6.0进行对比分析,采用N-J法构建新甲型H1N1流感病毒HA、NA基因进化树。结果廊坊市2009-2017年新甲型H1N1流感病毒HA基因序列同源性为97.2%~100.0%,NA基因序列同源性为97.7%~100.0%,HA和NA抗原决定簇、受体结合位点、糖基化位点均未发生变异。随着时间的推移,NA和HA突变位点均呈增多趋势,表明毒株逐年发生变异。结论 2009-2017年河北省廊坊市新甲型H1N1流感病毒HA和NA基因与国内、国际代表株高度同源,但其编码蛋白氨基酸的序列逐年发生变异,并出现不同程度的变异增强现象,因此应进一步加强对流感的监测。
Objective To investigate the epidemiology of a novel influenza A(H1N1 2009)virus isolated in Langfang,Hebei Province between 2009 and 2017 based on a phylogenetic analysis of the hemagglutinin(HA)and neuraminidase(NA)genes. Methods Fourteen strains of influenza A(H1N1 2009)were isolated by the Langfang CDC between 2009 and 2017.The sequences of the HA and NA genes of those strains were determined using PCR and sequencing.A phylogenetic tree was constructed for those strains using the N-J method and the software MEGA 6.0. Results The sequence of the HA gene of the novel influenza A(H1N1)virus was 97.2%-100.0% similar to that of the influenza A(H1N1)virus from 2009-2017.The sequence of the NA gene of the novel influenza A(H1N1)virus was 97.7%-100.0% similar to that of the influenza A(H1N1)virus from 2009-2017.There were no mutations in antigenic determinants,receptor binding sites,or glycosylation sites of the HA and NA genes.Results indicated that the number of mutations in the HA and NA genes correlated with the passage of time. Conclusion The current results indicated that influenza A(H1N1,2009)strains isolated between 2009 and 2017 in Langfang were similar to reference strains.This suggests that there were no large-scale outbreaks of H1N1 in Langfang during that period.However,H1N1 genes have continued to mutate,and surveillance needs be further enhanced in the future.
Objective To investigate the epidemiology of a novel influenza A(H1N1 2009)virus isolated in Langfang,Hebei Province between 2009 and 2017 based on a phylogenetic analysis of the hemagglutinin(HA)and neuraminidase(NA)genes. Methods Fourteen strains of influenza A(H1N1 2009)were isolated by the Langfang CDC between 2009 and 2017.The sequences of the HA and NA genes of those strains were determined using PCR and sequencing.A phylogenetic tree was constructed for those strains using the N-J method and the software MEGA 6.0. Results The sequence of the HA gene of the novel influenza A(H1N1)virus was 97.2%-100.0% similar to that of the influenza A(H1N1)virus from 2009-2017.The sequence of the NA gene of the novel influenza A(H1N1)virus was 97.7%-100.0% similar to that of the influenza A(H1N1)virus from 2009-2017.There were no mutations in antigenic determinants,receptor binding sites,or glycosylation sites of the HA and NA genes.Results indicated that the number of mutations in the HA and NA genes correlated with the passage of time. Conclusion The current results indicated that influenza A(H1N1,2009)strains isolated between 2009 and 2017 in Langfang were similar to reference strains.This suggests that there were no large-scale outbreaks of H1N1 in Langfang during that period.However,H1N1 genes have continued to mutate,and surveillance needs be further enhanced in the future.
引文
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[18]宋玥,颜文娟,方坤,等.2009至2016年中国甲型H1N1流感病毒神经氨酸酶基因进化分析及流行趋势预测[J].临床检验杂志,2017,35(7):550-4.
[19]章惠萍,易文晶,崔大伟.2013~2016年东部沿海地区甲型H1N1流感病毒分子特征分析[J].临床检验杂志,2017,35(4):306-9.
[20]秦剑秋,王亮,刘海燕,等.2013-2014年南宁市A(H3N2)亚型流感病毒病原学特征分析[J].中国病原生物学杂志,2017,12(6):560-5.
[21]颜文娟,郭艳,李小杉,等.2009-2015年中国甲型H1N1流感病毒血凝素基因进化分析[J].微生物学通报,2017,44(2):420-7.
[22] Ebranati E,Pariani E,Piralla A,et al.Reconstruction of the evolutionary dynamics of A(H3N2)influenza viruses circulating in Italy from 2004 to 2012[J].Plos One,2015,10(9):e0137099.
[2]孙新城,王云龙,李玉林,等.甲型流感H1N1(2009)流感病毒M2e单克隆抗体的制备及双抗体夹心ELISA检测方法的建立[J].中国病原生物学杂志,2015,10(6):508-11,524.
[3] Skehel JJ,Wiley DC.Receptor binding and membrane fusion in virus entry:the influenza hemagglutinin[J].Annu Rev Biochem,2000,69(1):531-69.
[4] Koel BF,Mogling R,Chutinimitkul S,et al.Identification of amino acid substitutions supporting antigenic change of influenza A(H1N1)pdm09viruses[J].J Virol,2015,89(7):3763-75.
[5] Gerlach T,Kühling L,Uhlendorff J,et al.Characterization of the neuraminidase of the H1N1/09pandemic influenza virus[J].Vaccine,2012,30(51):7348-52.
[6] Katz JM,Plowden J,Renshaw-Hoelscher M,et al.Immunity to influenza[J].J Infect Dis,1972,126(4):466-8.
[7] Nicholson KG,Wood JM,Zambon M.Influenza[J].Lancet,2003,362(9397):1733-45.
[8] Cyranoski D.Snapshot:trouble in paradise[J].Nature,2007,448(7155):738.
[9] Pariani E,Amendola A,Zappa A,et al.Molecular characterization of influenza viruses circulating in Northern Italy during two seasons(2005/2006and 2006/2007)of low influenza activity[J].J Med Virol,2008,80(11):1984-91.
[10] Webby RJ,Webster RG,Richt JA.Influenza viruses in animal wildlife populations.[J].Curr Top Microbiol Immun,2007,315(315):67.
[11]王晓亮,王静静,李知新,等.宁夏地区H1N1亚型猪流感病毒基因组遗传进化分析[J].中国兽医学报,2017,37(1):23-8.
[12]杨鹏飞,燕清丽,马雪征,等.聚集性感染甲型H1N1流感病毒的HA1基因特征分析[J].中华实验和临床病毒学杂志,2017,31(1):42-6.
[13] Zhang W,Qi J,Shi Y,et al.Crystal structure of the swine-origin A(H1N1)-2009influenza A virus hemagglutinin(HA)reveals similar antigenicity to that of the 1918pandemic virus[J].Protein Cell,2010,1(5):459-67.
[14] Sakabe S,Ozawa M,Takano R,et al.Mutations in PA,NP and HA of a pandemic(H1N1)2009influenza virus contribute to its adaptation to mice[J].Virus Res,2011,158(1):124-9.
[15] Al-Qahtani AA,Mubin M,Dela Cruz DM,et al.Phylogenetic and nucleotide sequence analysis of influenza A(H1N1)HA and NA genes of strains isolated from Saudi Arabia.[J].J Infect Dev Ctries,2017,11(1):81.
[16] Koo BS,Kim HK,Na W,et al.Complete genome sequence of an avian H1N1Influenza virus strain isolated from migratory birds in the Republic of Korea.[J].Genome Announ,2017,5(21):e00356.
[17]刘大飞,刘明,刘春国,等.H1N1亚型猪流感病毒广东分离株全基因克隆及其遗传演化分析[J].病毒学报,2008,24(5):358-63.
[18]宋玥,颜文娟,方坤,等.2009至2016年中国甲型H1N1流感病毒神经氨酸酶基因进化分析及流行趋势预测[J].临床检验杂志,2017,35(7):550-4.
[19]章惠萍,易文晶,崔大伟.2013~2016年东部沿海地区甲型H1N1流感病毒分子特征分析[J].临床检验杂志,2017,35(4):306-9.
[20]秦剑秋,王亮,刘海燕,等.2013-2014年南宁市A(H3N2)亚型流感病毒病原学特征分析[J].中国病原生物学杂志,2017,12(6):560-5.
[21]颜文娟,郭艳,李小杉,等.2009-2015年中国甲型H1N1流感病毒血凝素基因进化分析[J].微生物学通报,2017,44(2):420-7.
[22] Ebranati E,Pariani E,Piralla A,et al.Reconstruction of the evolutionary dynamics of A(H3N2)influenza viruses circulating in Italy from 2004 to 2012[J].Plos One,2015,10(9):e0137099.