2009—2018年广东省新甲型H1N1流感病毒奥司他韦耐药性分析

doi:10. 13217/j. scjpm. 2019. 0211

摘要/Abstract

摘要： 目的了解广东省2009—2018年分离到的新甲型H1N1[A（H1N1）pdm09]流感病毒对奥司他韦（商品名达菲）的耐药情况,为流感的临床治疗用药提供一定的指导,同时监测流感流行株的变异趋势。方法选取2009—2018年广东省网络实验室分离到的A（H1N1）pdm09流感病毒（每个网络实验室每年至少送检1~2株毒株）,用化学荧光为基础的中和抑制实验检测各病毒株的IC50值,筛选出奥司他韦的可疑耐药株;同时提取112株A（H1N1）pdm09流感病毒核酸,用RT-PCR方法扩增流感病毒的神经氨酸酶（NA）基因,通过病毒核酸测序并用生物信息学软件分析NA基因与耐药性有关的氨基酸位点及其基因变异情况。结果广东省分离到的A（H1N1）pdm09流感病毒有1.76%（5/284）对奥司他韦耐药,其H275Y位点（按N1编码序列）发生变异,即氨基酸275位点均由275H（组氨酸）变异为275Y（酪氨酸）,确定为奥司他韦的耐药株。 2009—2018年广东省分离的甲型H1N1毒株的NA基因按年份成簇分布,随着时间发展不断出现新的进化簇,具有一定的时段倾向性。结论 2009—2018年广东省的A（H1N1）pdm09流感病毒对奥司他韦的耐药比例低,奥司他韦对广东省的A（H1N1）pdm09流感病毒仍然敏感,病毒株的NA基因随时间发展不断出现新的进化簇,具有一定的时段倾向性,需继续开展抗流感病毒耐药性的监测,为临床抗流感病毒药物的使用提供科学依据。

关键词: 流感病毒A型, 奥司他韦, 耐药性, 基因

Abstract: Objective To explore the resistance of influenza A（H1N1）pdm09 isolated in Guangdong Province from 2009 to 2018 to oseltamivir （Tamiflu）, and monitor the variant trend of the epidemic strain of influenza virus. Methods Strains of influenza A（H1N1）pdm09 isolated from Guangdong Provincial Network Laboratories from 2009 to 2018 were selected （at least 1-2 strains per year in each network laboratory）.The IC50s of strains were detected by using fluorescence-based neutralization inhibition assay to select suspicious oseltamivir-resistant strains. At the same time, 112 strains of influenza A （H1N1） pdm09 virus nucleic acid were extracted and neuraminic acid （NA） gene of the virus was amplified by RT-PCR. Viral nucleic acid sequencing and bioinformatics software were used to analyze amino acid loci and gene mutations related to drug resistance of NA gene. Result Five strains（1.76%） were resistant to oseltamivir, had mutation in H275Y locus （according to N1 coding sequence）, i.e., amino acid locus 275 was all mutaed from 275H （histidine） to 275Y （tyrosine）, and then identified as Tamiflu resistant strains. The NA genes of A（H1N1）pdm09 strains isolated in Guangdong Province from 2009 to 2018 were clustered by year, and new evolutionary clusters appeared over time with a certain period of time. Conclusion Influenza A（H1N1）pdm09 virus has a very low resistance and still sensitive to oseltamivir in Guangdong Province. It is necessary to conduct surveillance of antiviral resistance of influenza virus continuously.

Key words: Influenza A virus, Oseltamivir, Resistance, Gene

中图分类号:

R183.3

刘丽儿, 邹丽容, 武婕, 宋颖超, 梁丽君, 郭前方, 庾健祥, 庄雪. 2009—2018年广东省新甲型H1N1流感病毒奥司他韦耐药性分析[J]. 华南预防医学, 2019, 45(3): 211-215.

LIU Li-er, ZOU Li-rong, WU Jie, SONG Ying-chao, LIANG Li-jun, GUO Qian-fang, YU Jian-xiang, ZHUANG Xue. Resistance of influenza A（H1N1）pdm09 to oseltamivir in Guangdong Province, China, 2009-2018[J]. South China Journal of Preventive Medicine, 2019, 45(3): 211-215.

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