2017年广州市16株H3N2流感病毒耐药基因分子特征分析
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摘要
目的分析广州市H3N2流感病毒耐药基因遗传进化和变异情况。方法对广州市2017年H3N2流感病毒进行分离,选择16株病毒对流感病毒耐药相关的NA(Neuraminidase,神经氨酸酶)和M2(Matrix protein 2,基质蛋白2)基因进行序列测定,使用DNA Star和MEGA软件对耐药基因的分子特征进行分析。结果本研究16株毒株在M2蛋白上均表现为对烷胺类药物耐药,NA蛋白的神经氨酸酶抑制剂作用位点均未发生突变,表现为对神经氨酸酶抑制剂敏感,但监测发现3C.2a.1分支内1株病毒在NA蛋白催化位点发生R224K变异。与疫苗株A/Hong Kong/4801/2014相比,16株毒株抗原位点和潜在糖基化位点都发生较大变异,抗原位点变异多发生在D339N,所有毒株在245位增加了一个糖基化位点NAT。基因进化分析发现,2017年广州市H3N2流感病毒呈现多分支进化特点,提示进化来源较为多样。结论监测发现3C.2a.1流行分支内出现酶活性位点变异毒株,因此需要重点关注变异株是否会随着3C.2a.1分支的流行而进一步扩散,以及酶活性位点的变异是否会降低病毒对神经氨酸酶抑制剂敏感性。
Objective To analyze the genetic evolution and variation of drug-resistance genes of influenza A(H3 N2) virus in Guangzhou.Methods Influenza A(H3 N2) viruses were isolated in Guangzhou in 2017.NA and M2 genes of influenza A(H3 N2) viruses from 16 strains were sequenced.DNA Star and MEGA software were used to perform sequence analysis.Results All strains in this study were resistant to alkanamines,but sensitive to neuraminidase inhibitors.Importantly,R224 K mutation was observed at NA protein of a strain of the virus in the 3 C.2 a.1 branch.Compared with vaccine strain-A/Hong Kong/4801/2014,variations at antigenic sites and glycosylation sites were observed,and the mutation of antigen mostly occurred at D339 N,and all strains added a glycosylation site NAT at 245.Gene evolution analysis showed that influenza A(H3 N2) viruses in Guangzhou exhibited the characteristic of multi-branch evolution,suggesting that the sources of evolution were diverse.Conclusion There is a mutant strain at enzyme active sites in the 3 C.2 a.1 branch.Therefore,we need to focus on whether the mutation will spread along with the prevalence of 3 C.2 a.1 strains and whether the mutation of enzyme active sites can reduce the susceptibility of influenza A(H3 N2) virus to neuraminidase inhibitors.
Objective To analyze the genetic evolution and variation of drug-resistance genes of influenza A(H3 N2) virus in Guangzhou.Methods Influenza A(H3 N2) viruses were isolated in Guangzhou in 2017.NA and M2 genes of influenza A(H3 N2) viruses from 16 strains were sequenced.DNA Star and MEGA software were used to perform sequence analysis.Results All strains in this study were resistant to alkanamines,but sensitive to neuraminidase inhibitors.Importantly,R224 K mutation was observed at NA protein of a strain of the virus in the 3 C.2 a.1 branch.Compared with vaccine strain-A/Hong Kong/4801/2014,variations at antigenic sites and glycosylation sites were observed,and the mutation of antigen mostly occurred at D339 N,and all strains added a glycosylation site NAT at 245.Gene evolution analysis showed that influenza A(H3 N2) viruses in Guangzhou exhibited the characteristic of multi-branch evolution,suggesting that the sources of evolution were diverse.Conclusion There is a mutant strain at enzyme active sites in the 3 C.2 a.1 branch.Therefore,we need to focus on whether the mutation will spread along with the prevalence of 3 C.2 a.1 strains and whether the mutation of enzyme active sites can reduce the susceptibility of influenza A(H3 N2) virus to neuraminidase inhibitors.
引文
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[3] Cui D,Zhao D,Xie G,et al.Simultaneous detection of influenza A subtypes of H3N2 virus,pandemic (H1N1) 2009 virus and reassortant avian H7N9 virus in humans by multiplex one-step real-time RTPCR assay[J].SpringerPlus,2016,5:2054.
[4] 鄢翠林,崔大伟.2016-2017年我国甲型流感病毒H3N2亚型HA和NA基因的分子特征分析[J].临床检验杂志,2018,36(1):66-69.
[5] 陈国清,李春香,邵荣标,等.2015-2017年盐城市甲型H3N2 流感病毒HA1、NA 基因分子特征[J].中华疾病控制杂志,2018,22(9):902-907.
[6] 赵升,僧明华,吕宛玉,等.2013-2017年河南省H3N2亚型流感病毒流行特征分析[J].现代预防医学,2018,45(7):1180-1183.
[7] 焦素黎,王蓉,倪红霞,等.浙江省宁波市2014～2015年H3N2流行性感冒病毒HA1基因与NA基因特性分析[J].中国疫苗和免疫,2016,22(3):312-316.
[8] 孙海波,刘双,王璐璐,等.辽宁省2014年-2016年季节性A(H3N2)流感病毒耐药基因分析[J].中国卫生检验杂志,2018,28(2):157-159.
[9] 苏文哲,柯昌文.流行性感冒病毒的耐药性相关研究进展[J].中国病毒病杂志,2011,1(3):171-176.
[10] 秦剑秋,裴建新,林新勤,等.南宁分离株A(H3N2)流感病毒神经氨酸酶遗传变异及蛋白结构分析[J].中国病原生物学杂志,2015,10(7):577-581,586.
[11] 梁丽君,黄平,侯年妹,等.广东流感H3N2 亚型神经氨酸酶基因变异及耐药分析[J].中华临床医师杂志(电子版),2012,6(6):1447-1451.
[12] 黄维娟,谭敏菊,李希妍,等.2013-2017年中国大陆流行的A(H3N2)亚型流感病毒对神经氨酸酶抑制剂的敏感性分析[J].病毒学报,2018,34(6):793-799.
[13] Colman PM,Varghese JN,Laver WG.Structure of the catalytic and antigenic sites in influenza virus neuraminidase[J].Nature,1983,303(5912):41-44.
[14] Zhong J,Liang L,Huang P,et al.Genetic mutations in influenza H3N2 viruses from a 2012 epidemic in Southern China[J].Virology Journal,2013,10(1):345.
[15] Rambaut A,Pybus OG,Nelson MI,et al.The genomic and epidemiological dynamics of human influenza A virus[J].Nature,2008,453(7195):615-619.