南京地区诺如病毒GⅡ.Pe-GⅡ.4基因特征
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摘要
目的研究南京地区诺如病毒GⅡ.Pe-GⅡ.4基因特征和变化规律,了解其进化过程及高变区氨基酸变化情况,预测进化方向及流行趋势,为疫情预警和处置提供科学依据。方法应用荧光定量PCR和一步法RT-PCR,对南京地区2014—2017年分离的10株GⅡ.Pe-GⅡ.4的部分聚合酶—衣壳蛋白区及P2区进行测序,并利用分子生物学软件对序列进行比对分析。结果进化树分析发现,2014—2015年和2016—2017年南京流行株分别位于两个簇。氨基酸分析显示,10株南京流行株同源性达97.3%～100.0%,与21株国内外2011—2017年参考株同源性达97.1%～100.0%。与2008年前驱期澳洲株(KX789174)P2区相比较,10株南京株共19个氨基酸位点发生改变,其中10个位点位于5个抗原表位。南京株2016年后抗原表位A区有氨基酸位点改变。结论 2014—2017年南京地区分离的诺如病毒GⅡ.Pe-GⅡ.4基因型流行株氨基酸同源性较高,但仍呈现出随时间迁移某些抗原位点的氨基酸改变,应持续观察其进化趋势。
Objective To study the genetic characteristics and changes of Norovirus GⅡ.Pe-GⅡ.4 in Nanjing;to understand the evolution process and amino acid changes in high-variant regions;to predict virus evolution direction and epidemic trend;to provide scientific basis for warning and treatment.Methods The fluorescence quantitative PCR and one-step RT-PCR were used to perform sequencing analysis for polymerase-capsid protein region and P2 region of 10 GⅡ.Pe-GⅡ.4 strains isolated in Nanjing from 2014 to 2017;the sequences were compared and analyzed by molecular biology software.Results The phylogenetic analysis revealed that Nanjing epidemic strains were categorized into 2 clusters in 2014-2015 and 2016-2017 epidemics,respectively.Amino acid analysis showed that the homology of 10 Nanjing epidemic strains was 97.3%-100.0%,and 97.1%-100.0% homologous to 21 domestic and foreign reference strains from 2011 to 2017.Compared with the P2 region of the 2008 precursor Australian strain(KX789174),19 amino acid loci were altered in 10 Nanjing strains,10 of which were located in 5 antigenic epitopes.The amino acid loci of antigen epitope A of Nanjing strains changed since 2016.Conclusion From 2014 to2017,the amino acid homology of the prevalent strains of Norovirus GⅡ.Pe-GⅡ.4 genotype isolated in Nanjing is relatively high;however the amino acid changes of some antigenic sites upon time.The virus evolutionary trend should be continuously monitored.
Objective To study the genetic characteristics and changes of Norovirus GⅡ.Pe-GⅡ.4 in Nanjing;to understand the evolution process and amino acid changes in high-variant regions;to predict virus evolution direction and epidemic trend;to provide scientific basis for warning and treatment.Methods The fluorescence quantitative PCR and one-step RT-PCR were used to perform sequencing analysis for polymerase-capsid protein region and P2 region of 10 GⅡ.Pe-GⅡ.4 strains isolated in Nanjing from 2014 to 2017;the sequences were compared and analyzed by molecular biology software.Results The phylogenetic analysis revealed that Nanjing epidemic strains were categorized into 2 clusters in 2014-2015 and 2016-2017 epidemics,respectively.Amino acid analysis showed that the homology of 10 Nanjing epidemic strains was 97.3%-100.0%,and 97.1%-100.0% homologous to 21 domestic and foreign reference strains from 2011 to 2017.Compared with the P2 region of the 2008 precursor Australian strain(KX789174),19 amino acid loci were altered in 10 Nanjing strains,10 of which were located in 5 antigenic epitopes.The amino acid loci of antigen epitope A of Nanjing strains changed since 2016.Conclusion From 2014 to2017,the amino acid homology of the prevalent strains of Norovirus GⅡ.Pe-GⅡ.4 genotype isolated in Nanjing is relatively high;however the amino acid changes of some antigenic sites upon time.The virus evolutionary trend should be continuously monitored.
引文
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[2] Aliabadi N,Lopman BA,Parashar UD,et al.Progress toward norovirus vaccines:considerations for further development and implementation in potential target populations[J].Expert Rev Vaccines,2015,14(9):1241-1253.
[3] Vinje,J.Advances in laboratory methods for detection and typing of norovirus[J].J Clin Microbiol,2015,53(2):373-381.
[4] Siebenga JJ, Lemey P, Kosakovsky Pond SL, et al.Phylodynamic reconstruction reveals norovirus GII.4epidemic expansions and their molecular determinants[J].PLoS Pathog,2010,6(5):e1000884.
[5] Van Beek J,Ambert-Balay K,Botteldoorn N,et al.Indications for worldwide increased norovirus activity associated with emergence of a new variant of genotype II.4,late 2012[J].Euro Surveill,2013,18(1):8-9.
[6] Shen Z,Wang G,Zai SB,et al.The emergence of novel GII.4norovirus variant,Sydney_2012,in Shanghai,China[J].Bing Du Xue Bao,2013,29(6):608-614.
[7] Emergence of new norovirus strain GII.4Sydney-United States,2012[J].MMWR,2013,62(3):55.
[8] Bennett S,MacLean A,Miller RS,et al.Increased norovirus activity in Scotland in 2012is associated with the emergence of a new norovirus GII.4variant[J].Euro Surveill,2013,18(2):1-2.
[9] Bruggink LD,Moselen JM,Roberts JA,et al.Evolutionary changes in the capsid P2region of Australian strains of the norovirus GII.Pe_GII.4[J].J Med Microbiol,2017,66(7):1014-1022.
[10] Wang X,Du X,Yong W,et al.Genetic characterization of emergent GII.17 norovirus variants from 2013 to 2015 in Nanjing,China[J].J Med Microbiol,2016,65(11):1274-1280.
[11]王璇,雍玮,杜雪飞,等.2014—2017年南京地区胃肠炎暴发中诺如病毒分子流行病学特征[J].病毒学报,2018,34(4):483-490.
[12] Donaldson EF,Lindesmith LC,Lobue AD,et al.Norovirus pathogenesis:mechanisms of persistence and immune evasion in human populations[J].Immunol Rev,2008,225:190-211.
[13] Lindesmith LC,Beltramello M,Donaldson EF,et al.Immunogenetic mechanisms driving norovirus GII.4antigenic variation[J].LoS Pathog,2012,8(5):e1002705.