2017-2018年广州市甲型H1N1流感病毒耐药基因变异监测
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摘要
目的对广州市流感监测人群呼吸道标本进行甲型H1N1流感病毒分离和测序,并对耐药基因突变和遗传进化特征进行分析。方法 2017年1月-2018年3月,对4家流感监测医院的5 831份监测标本进行甲型H1N1流感病毒核酸检测,将阳性标本接种MDCK细胞进行流感病毒分离和测序,应用DNA Star7.1软件和MEGA 4.0软件对耐药基因进行测序分析。结果 5 831份监测标本中检出甲型H1N1核酸阳性标本222份,检出率为3.81%,其中分离甲型H1N1流感病毒61株,分离率为1.05%。有2株病毒分离株出现了H274Y突变,突变率为3.28%。所有病毒分离株均发生了S31N突变,突变率为100%,同时另有2株出现V27A突变,突变率为3.28%。部分2017年毒株和2018年毒株已形成于A/Michigan/45/2015疫苗株分支之外的另一独立小分支。结论 2017年广州市流感监测病例中监测到2株H1N1流感病毒分离株为奥司他韦耐药突变株,且该耐药株均位于当前疫苗株流行分支以外的独立分支,有必要对广州市甲型H1N1流感病毒进行持续监测。
Objective To isolate and sequence H1 N1 influenza virus isolated from respiratory specimens of influenza surveillance population in Guangzhou,and to analyze the mutation and evolution characteristics of drug resistance genes.Methods From January 2017 to March,1818,5,831 surveillance samples from 4 influenza surveillance hospitals were tested for H1 N1 influenza virus nucleic acid.Positive samples were inoculated into MDCK cells for influenza virus isolation and sequencing.DNA Star7.1 software was applied.The drug resistance gene was sequenced and analyzed with MEGA 4.0 software.Results A total of 222 H1 N1 positive samples were detected in the respiratory tract specimens of 5 831 influenza-like cases,with a detection rate of 3.81%.61 strains of A H1 N1 influenza virus were isolated.There were 2 isolates with H274 Y mutation in NA gene.All strains had S31 N mutation in M2 gene.Phylogenetic analysis showed that some isolates from 2017 and 2018 were different from the A/Michigan/45/2015 branch.Conclusion In 2017,two A H1 N1 influenza strains with H274 Y mutation in NA gene were detected in influenza surveillance cases in Guangzhou for the first time,which belonged to other prevalent branches different from the current vaccine strains.Therefore,it is necessary to continuously monitor the mutation and evolution of the resistance genes A H1 N1 influenza virus in Guangzhou.
Objective To isolate and sequence H1 N1 influenza virus isolated from respiratory specimens of influenza surveillance population in Guangzhou,and to analyze the mutation and evolution characteristics of drug resistance genes.Methods From January 2017 to March,1818,5,831 surveillance samples from 4 influenza surveillance hospitals were tested for H1 N1 influenza virus nucleic acid.Positive samples were inoculated into MDCK cells for influenza virus isolation and sequencing.DNA Star7.1 software was applied.The drug resistance gene was sequenced and analyzed with MEGA 4.0 software.Results A total of 222 H1 N1 positive samples were detected in the respiratory tract specimens of 5 831 influenza-like cases,with a detection rate of 3.81%.61 strains of A H1 N1 influenza virus were isolated.There were 2 isolates with H274 Y mutation in NA gene.All strains had S31 N mutation in M2 gene.Phylogenetic analysis showed that some isolates from 2017 and 2018 were different from the A/Michigan/45/2015 branch.Conclusion In 2017,two A H1 N1 influenza strains with H274 Y mutation in NA gene were detected in influenza surveillance cases in Guangzhou for the first time,which belonged to other prevalent branches different from the current vaccine strains.Therefore,it is necessary to continuously monitor the mutation and evolution of the resistance genes A H1 N1 influenza virus in Guangzhou.
引文
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[13] 付倩,文奇,周银古,等.新余市甲型流感病毒M基因进化及金刚烷胺类耐药性分析[J].实验与检验医学,2017,35(6):870-873.
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[15] 苏彤,李淑华,鹿文英,等.一株新型甲型H1N1流感病毒H275Y的奥司他韦耐药变异分析[J].解放军医学杂志,2009,34(12):1412-1418.
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[2] Wu Y,Wu Y,Tefsen B,et al.Bat-derived influenza-like viruses H17N10 and H18N11[J].Trends in Microbiology,2014,22(4):183-191.
[3] Webster RG,Bean WJ,Gorman OT,et al.Evolution and ecology of influenza A viruses[J].Microbiological Reviews,1992,56(1):152-179.
[4] Fouchier MR,Munster V,Wallensten A,et al.Characterization of a novel influenza A virus hemagglutinin subtype(HI6)obtained from black-headed gulls[J].Journal of Virology,2005,79(5):2814-2822.
[5] Knipe DM,Howley PM,Griffin DE,et al.Fields virology[M].5th ed.Philadelphia:Lippincott Williams and Wilkins,2007:1647-1689.
[6] 王昕,房师松,彭博,等.深圳市急性呼吸道感染住院和门诊病例的流感流行特征分析[J].中国卫生检验杂志,2016,26(1):107-108,112.
[7] 许沙沙,常彦敏,徐霖,等.2010年广州市甲型H1N1流感病毒分离株NA基因变异分析[J].检验医学,2014,29(1):42-49.
[8] 郭雪,马燕丽,康艳先,等.流感病毒裂解疫苗接种人体后的安全性和免疫原性研究[J].中华微生物学和免疫学杂志,2018,38(5):361-365.
[9] Pinto LH,Lamb RA.Understanding the mechanism of action of the anti-influenza virus drug amantadine[J].Trends in Microbiology,1995,3(7):271.
[10] 谢剑锋,张炎华,赵琳,等.一株达菲耐药的甲型H1N1流感病毒全基因特征分析[J].中国微生态学杂志,2016,28(8):884-888.
[11] 赵丹,苏志磊,于萍,等.青岛地区2012-2014年甲型H1N1流感病毒基因特征[J].中国公共卫生,2016,32(8):1096-1100.
[12] 许爽,杨显达,吴东林,等.16株甲型流感病毒耐药性基因测序结果分析[J].中国卫生工程学,2010,9(1):54-55.
[13] 付倩,文奇,周银古,等.新余市甲型流感病毒M基因进化及金刚烷胺类耐药性分析[J].实验与检验医学,2017,35(6):870-873.
[14] 熊英,龚甜,周珺,等.江西省2009-2011年新型甲型H1N1流感病毒耐药性分析[J].现代预防医学,2014,41(9):1681-1685,1688.
[15] 苏彤,李淑华,鹿文英,等.一株新型甲型H1N1流感病毒H275Y的奥司他韦耐药变异分析[J].解放军医学杂志,2009,34(12):1412-1418.
[16] 郭元吉,程小雯.流行性感冒病毒及其实验技术[M].北京:中国三峡出版社,1997.