Assessment of Pandemic and Seasonal Influenza A (H1N1) Virus Susceptibility to Neuraminidase Inhibitors in Three Enzyme Activity Inhibition Assays
Assessment of Pandemic and Seasonal Influenza A (H1N1) Virus Susceptibility to Neuraminidase Inhibitors in Three Enzyme Activity Inhibition Assays
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DOI：
10.1128/aac.00581-10
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发表时间：
2010-09-01
影响因子：
4.9
通讯作者：
Gubareva, Larisa V.
中科院分区：
文献类型：
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作者：
Nguyen, Ha T.;Sheu, Tiffany G.;Gubareva, Larisa V.
神经氨酸酶抑制剂 (NAIs) 扎那米韦和奥司他韦是目前唯一能有效治疗和预防 2009 年大流行甲型流感 (H1N1) 病毒感染的抗病毒药物。已证实这些病毒在治疗期间获得 NAI 耐药性的潜力强调了评估其 NAI 敏感性的必要性。已知 50% 抑制浓度 (IC(50)) 会根据所使用的神经氨酸酶抑制 (NI) 测试而变化；然而，很少对不同的 NI 检测进行并排比较。在本研究中，代表 2009 年季节性和大流行性 H1N1 流感病毒（包括奥司他韦耐药 H275Y 变种）的 11 个分离株在三种功能 NI 测定中进行了测试：化学发光 (CL)、荧光 (FL) 和比色 (CM)。评估了病毒对扎那米韦、奥司他韦和三种研究性 NAI（帕拉米韦、R-125489 和 A-315675）的敏感性。通过所有三种 NI 测定，除 H275Y 变体外，所有分离株对所有五种 NAI 均敏感。 H275Y 变体显示出针对奥司他韦和帕拉米韦的 IC(50) 显着升高。三种 NI 检测总体上得出一致的结果；因此，NI 测定的选择似乎不会影响基于药物敏感性监测的结论。然而，与其他测定相比，每种测定都具有一定的优势：CL 测定需要较少的病毒体积，FL 测定提供了野生型和变体之间 IC(50) 的最大差异，而从 CM 测定获得的 IC(50) 可能最能预测抑制人类酶活性所需的药物浓度。期望开发一种NI测定法，其结合了所有三种当前可用测定法的优点但缺乏它们的缺点。
The neuraminidase inhibitors (NAIs) zanamivir and oseltamivir are currently the only antiviral drugs effective for the treatment and prophylaxis of 2009 pandemic influenza A (H1N1) virus infections. The proven potential of these viruses to acquire NAI resistance during treatment emphasizes the need to assess their NAI susceptibility. The 50% inhibitory concentrations (IC(50)s) are known to vary depending on the neuraminidase inhibition (NI) test used; however, few side-by-side comparisons of different NI assays have been done. In the present study, a panel of 11 isolates representing 2009 seasonal and pandemic influenza H1N1 viruses, including oseltamivir-resistant H275Y variants, were tested in three functional NI assays: chemiluminescent (CL), fluorescent (FL), and colorimetric (CM). The sensitivities of the viruses to zanamivir, oseltamivir, and three investigational NAIs (peramivir, R-125489, and A-315675) were assessed. All isolates with the exception of H275Y variants were sensitive to all five NAIs by all three NI assays. The H275Y variants showed substantially elevated IC(50)s against oseltamivir and peramivir. The three NI assays generally yielded consistent results; thus, the choice of NI assay does not appear to affect conclusions based on drug susceptibility surveillance. Each assay, however, offers certain advantages compared to the others: the CL assay required less virus volume and the FL assay provided the greatest difference in the IC(50)s between the wild type and the variants, whereas the IC(50)s obtained from the CM assay may be the most predictive of the drug concentrations needed to inhibit enzyme activity in humans. It would be desirable to develop an NI assay which combines the advantages of all three currently available assays but which lacks their shortcomings.