Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta Georgia 30333, USA.
Antimicrob Agents Chemother. 2010 Mar;54(3):1102-10. doi: 10.1128/AAC.01417-09. Epub 2009 Dec 22.
The M2 blockers amantadine and rimantadine and the neuraminidase (NA) inhibitors (NAIs) oseltamivir and zanamivir are approved by the FDA for use for the control of influenza A virus infections. The 2009 pandemic influenza A (H1N1) viruses (H1N1pdm) are reassortants that acquired M and NA gene segments from a Eurasian adamantane-resistant swine influenza virus. NAI resistance in the H1N1pdm viruses has been rare, and its occurrence is mainly limited to oseltamivir-exposed patients. The pyrosequencing assay has been proven to be a useful tool in surveillance for drug resistance in seasonal influenza A viruses. We provide a protocol which allows the detection of adamantane resistance markers as well as the I43T change, which is unique to the H1N1pdm M2 protein. The protocol also allows the detection of changes at residues V116, I117, E119, Q136, K150, D151, D199, I223, H275, and N295 in the NA, known to alter NAI drug susceptibility. We report on the detection of the first cases of the oseltamivir resistance-conferring mutation H275Y and the I223V change in viruses from the United States using the approach described in this study. Moreover, the assay permits the quick identification of the major NA group (V106/N248, I106/D248, or I106/N248) to which a pandemic virus belongs. Pyrosequencing is well suited for the detection of drug resistance markers and signature mutations in the M and NA gene segments of the pandemic H1N1 influenza viruses.
M2 阻滞药金刚烷胺和金刚乙胺以及神经氨酸酶 (NA) 抑制剂 (NAIs) 奥司他韦和扎那米韦已获得美国食品药品监督管理局批准，可用于控制甲型流感病毒感染。2009 年大流行性流感 A 型 (H1N1) 病毒 (H1N1pdm) 是重配病毒，其获得了欧亚地区抗金刚烷胺猪流感病毒的 M 和 NA 基因片段。H1N1pdm 病毒中的 NAI 耐药性很少见，主要发生在接受奥司他韦暴露的患者中。焦磷酸测序检测已被证明是监测季节性甲型流感病毒耐药性的有用工具。我们提供了一个检测方案，该方案可检测到金刚烷胺耐药性标记物以及 H1N1pdm M2 蛋白所特有的 I43T 变化。该方案还可检测到 NA 中已知改变 NAI 药物敏感性的残基 V116、I117、E119、Q136、K150、D151、D199、I223、H275 和 N295 的变化。我们报告了使用本研究中描述的方法在美国检测到的首例奥司他韦耐药性赋予突变 H275Y 和 I223V 变化的病毒。此外，该检测方法还可快速确定大流行病毒所属的主要 NA 组 (V106/N248、I106/D248 或 I106/N248)。焦磷酸测序非常适合检测大流行性甲型流感病毒 M 和 NA 基因片段中的耐药性标记物和特征性突变。
