广东地区新甲型H1N1流感病毒对奥司他韦的耐药性分析  

Surveillance for Oseltamivir-resistant influenza pandemic A H1N1 viruses in Guangdong province

作　　者：邹丽容[1] 周杰[1] 管大伟[1] 张欣[1] 倪汉忠[1] 黄平[1] 武婕[1] 李晖[1] 柯昌文[1] 

机构地区：[1]广东省疾病预防控制中心病原微生物检验所WHO新发传染病监测培训合作中心广东省应急病原学重点实验室,广州510300

出　　处：《中华检验医学杂志》2011年第10期940-944,共5页Chinese Journal of Laboratory Medicine

基　　金：广东省医学科学技术研究基金资助项目（A2010066）

摘　　要：目的 调查广东省2009年新甲型H1N1流感病毒对奥司他韦的耐药情况,为临床用药提供指导,监测流感流行株的变异趋势.方法 2009年4-12月,对广东省流感监测哨点医院收集患者鼻咽拭子,用MDCK细胞或者鸡胚,分离得到流感毒株221株.利用神经氨酸酶化学荧光抑制实验检测毒株对奥司他韦的IC50,筛选出对神经氨酸酶抑制剂的敏感性下降的毒株.同时选取68株病毒,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 分离到新甲型H1N1流感病毒221株全部对奥司他韦敏感,IC50中位数为0.24 nmol/L,最小值为0.02 nmol/L,最大值为1.66 nmol/L.NA测序结果也没有发现导致耐药的突变位点.进化分析表明,广东省的新甲型H1N1流感病毒的NA基因与全球流行的新甲型H1N1流感病毒的核苷酸同源性在99.5％～100.0％之间.结论 奥司他韦对广东地区的新甲型H1N1流感病毒有效,可以继续用来预防和治疗流感患者.新甲型H1N1流感病毒的NA基因变化不大.Objective To investigate the susceptibility to oseltamivir of influenza pandemic A （H1N1） 2009 viruses in Guangdong province during 2009,provide valuable information of prescription for clinics,and elucidate the variant trend of the epidemic strain based on phylogenetic analysis.Methods During April to December 2009,clinical specimens were collected from sentinel hospitals covering the whole Guangdong province.Virus isolation was performed by in MDCK cells or embryonated chicken eggs.A fluorescence-based neuraminidase （NA） enzyme inhibition assay was conducted to measure influenza susceptibility.The NAI susceptibility of influenza virus was expressed as the concentration of NAI needed to inhibit the NA enzyme activity by 50%.A subset of 68 viruses were performed NA sequencing for detecting resistant mutations and studying variant trends.Results During surveillance,221 influenza pandemic A （ H1N1 ） viruses were isolated.All strains were sensitive to oseltamivir inhibition assay,with a median IC50 of 0.24 nmol/L （range 0.02 -1.66 nmol/L）.No mutation related to resistance was found.Phylogenic analysis illustrated that these NA genes were homology high to 99.5% - 100.0% with those from other countries.Conclusions influenza pandemic A （H1N1） 2009 viruses were sensitive to oseltamivir in Guangdong,and useful for prophylaxis and treatment of influenza infection.Little selective pressure was found by phylogenic analysis.Our laboratory will continue to observe antiviral-resistance among circulating influenza viruses.

关 键 词：流感病毒A型 H1N1亚型 奥司他韦 抗药性 病毒 

分 类 号：R51[医药卫生—内科学]