2017-2018年山东省甲型H1N1流感病毒耐药性分析
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摘要
目的调查山东省2017-2018流感监测年度甲型H1N1流感病毒对神经氨酸酶抑制剂(NAI)的耐药情况,分析其神经氨酸酶(NA)基因特征。方法选取山东省2017-2018流感监测年度分离的20株甲型H1N1流感病毒进行生物学耐药实验,检测病毒对奥司他韦和扎那米韦的药物敏感性。提取病毒核酸后对NA基因进行测序,利用生物信息学软件分析NA基因上与耐药有关的氨基酸位点及其基因变异情况。结果选取的20株甲型H1N1流感病毒全部对奥司他韦和扎那米韦敏感,对奥司他韦的IC50平均数为0.37 n M(0.15～0.89 n M),对扎那米韦的IC50平均数为0.28 n M(0.14～0.78 n M)。NA基因测序结果也没有发现导致耐药的突变位点。结论山东省的甲型H1N1流感病毒对NAI敏感,临床上可继续使用此类药物对流感患者进行治疗。
Objective To investigate the susceptibility of influenza pandemic A(H1 N1) viruses to neuraminidase inhibitors,and to analyze the molecular characteristics of neuraminidase(NA) genes in Shandong Province from 2017 to 2018 influenza surveillance year. Methods Antiviral resistant phenotyping test was performed to analyze the NAI susceptibility of 20 influenza pandemic A(H1 N1) viruses isolated in Shandong Province from 2017 to 2018, including the test of susceptibility to oseltamivir and zanamivir. NA genes were sequenced after viral RNA was extracted. The nucleic and amino acid sequences were analyzed to detect the resistant mutations and study the variant trends. Results All 20-influenza pandemic A(H1 N1)viruses tested were sensitive to oseltamivir and zanamivir. The median IC50 of the viruses for oseltamivir was of 0.37 nM(range 0.15-0.89 nM) while 0.28 nM(range 0.14-0.78 nM) for zanamivir.No mutation related to medicine resistance was found. Conclusion Influenza pandemic A(H1 N1) 2009 viruses were sensitive to NAIs in Shandong which indicated that NAIs could be used continually for clinical treatment of patients with influenza.
Objective To investigate the susceptibility of influenza pandemic A(H1 N1) viruses to neuraminidase inhibitors,and to analyze the molecular characteristics of neuraminidase(NA) genes in Shandong Province from 2017 to 2018 influenza surveillance year. Methods Antiviral resistant phenotyping test was performed to analyze the NAI susceptibility of 20 influenza pandemic A(H1 N1) viruses isolated in Shandong Province from 2017 to 2018, including the test of susceptibility to oseltamivir and zanamivir. NA genes were sequenced after viral RNA was extracted. The nucleic and amino acid sequences were analyzed to detect the resistant mutations and study the variant trends. Results All 20-influenza pandemic A(H1 N1)viruses tested were sensitive to oseltamivir and zanamivir. The median IC50 of the viruses for oseltamivir was of 0.37 nM(range 0.15-0.89 nM) while 0.28 nM(range 0.14-0.78 nM) for zanamivir.No mutation related to medicine resistance was found. Conclusion Influenza pandemic A(H1 N1) 2009 viruses were sensitive to NAIs in Shandong which indicated that NAIs could be used continually for clinical treatment of patients with influenza.
引文
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[11] Abe Y, Takashita E, Sugawara K, et al. Effect of the addition ofoligosaccharides on the biological activities and antigenicity of in-fluenza A/H3N2 virus hemagglutinin[J]. Journal of Virology, 2004,78(18):9605-9611.
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[14]田疆,周经姣,陈艺韵,等.甲型H1N1流感病毒神经氨酸酶基因遗传进化分析[J].中山大学学报(医学科学版),2010,2(2):207-212.
[15]黄平,俞守义,柯昌文.逐步预测和统计学筛选人H_5N_1毒株神经氨酸酶蛋白B细胞表位[J].科学通报,2008,(22):2748-2753.
[2] Novel Swine-Origin Influenza A(H1N1)Virus Investigation Team,Dawood FS, Jain S, et al. Emergence of a novel swine-origin in-fluenza A(H1N1)virus in humans[J]. The New England Journal ofMedicine, 2009, 360(25):2605-2615.
[3] Nguyen HT, Sheu TG, Mishin VP, et al. Assessment of pandemic andseasonal influenza A(H1N1)virus susceptibility to neuraminidaseinhibitors in three enzyme activity inhibition assays[J]. AntimicrobialAgents and Chemotherapy, 2010, 54(9):3671-3677.
[4] Okomo-Adhiambo M, Sleeman K, Ballenger K, et al.Neuraminidaseinhibitor susceptibility testing in human influenza viruses:a labora-tory surveillance perspective[J]. Viruses, 2010, 2(10):2269-2289.
[5] Meetings of the WHO working group on surveillance of influenza an-tiviral susceptibility-Geneva, November 2011 and June 2012[J].Releve Epidemiologique Hebdomadaire, 2012, 87(39):369-374.
[6] Update on oseltamivir-resistant pandemic A(H1N1)2009 influen-za virus:January 2010[J]. Releve Epidemiologique Hebdomadaire,2009, 85(6):37-40.
[7]谢剑锋,沈晓娜,王美爱,等.福建省甲型H1N1流感病毒分离及首例分离株全基因组序列分析[J].中国人兽共患病学报,2009,(8):745-748, 809.
[8]李娟,赵晓南,曹艺会,等.云南省2009~2014年甲型H1N1流感病毒HA及NA基因特征分析[J].病毒学报,2015,(6):674-678.
[9] Zhou J, Zou L, Zhang X, et al. Adamantane-and oseltamivir-re-sistant seasonal A(H1N1)and pandemic A(H1N1)2009 influenzaviruses in Guangdong, China, during 2008 and 2009[J]. Journal ofClinical Microbiology, 2011, 49(7):2651-2655.
[10]吕星,吴春利,阳帆,等.深圳市2008-2009年A型H1N1季节性流感病毒神经氨酸酶抑制剂耐药性监测[J].中华微生物学和免疫学杂志,2011,31(7):609-612.
[11] Abe Y, Takashita E, Sugawara K, et al. Effect of the addition ofoligosaccharides on the biological activities and antigenicity of in-fluenza A/H3N2 virus hemagglutinin[J]. Journal of Virology, 2004,78(18):9605-9611.
[12] Schulze IT. Effects of glycosylation on the properties and functions ofinfluenza virus hemagglutinin[J]. The Journal of Infectious Diseases,1997, 176(Suppl 1):S24-S28.
[13] Colman PM, Varghese JN, Laver WG. Structure of the catalytic andantigenic sites in influenza virus neuraminidase[J]. Nature, 1983,303(5912):41-44.
[14]田疆,周经姣,陈艺韵,等.甲型H1N1流感病毒神经氨酸酶基因遗传进化分析[J].中山大学学报(医学科学版),2010,2(2):207-212.
[15]黄平,俞守义,柯昌文.逐步预测和统计学筛选人H_5N_1毒株神经氨酸酶蛋白B细胞表位[J].科学通报,2008,(22):2748-2753.