Dr. Christophe Mérieux Laboratory, IPB, CAMS-Fondation Mérieux and State Key Laboratory of Molecular Virology and Genetic Engineering, Institute of Pathogen Biology, Chinese Academy of Medical Sciences, Beijing 100730, China.
J Virol Methods. 2010 Jul;167(1):37-44. doi: 10.1016/j.jviromet.2010.03.007. Epub 2010 Mar 19.
Pandemic influenza A/H1N1 2009 and seasonal influenza viruses are currently co-circulating worldwide. A rapid, sensitive, and specific assay for distinguishing pandemic influenza A/H1N1 2009 from seasonal influenza viruses and for subtyping seasonal influenza A viruses could aid in the surveillance and control of these viral infections. Here, such a multiplex real-time RT-PCR (rRT-PCR) assay for typing influenza A and B viruses and the pandemic influenza A/H1N1 2009 is developed. This assay can also subtype seasonal influenza A viruses simultaneously. The analytical sensitivity is 10-10(4) copies/reaction. The coefficients of variation of inter-assay and intra-assay are 0.04-0.45% and 0.08-0.97%, respectively. The new multiplex rRT-PCR assay is more sensitive in subtyping seasonal influenza viruses than the conventional PCR techniques. Results obtained with this assay for the detection of pandemic influenza A/H1N1 2009 are highly consistent (96.88%) with those achieved using the US CDC's rRT-PCR protocol. A sample identified as "pandemic influenza A/H1N1 2009 positive" by the US CDC's rRT-PCR was reclassified correctly as subtype H3N2 using this assay. Taken together, this new multiplex rRT-PCR protocol could be an important tool for improving diagnosis and management of the pandemic influenza A/H1N1 2009 and seasonal influenza viruses.
目前，大流行流感 A/H1N1 2009 病毒和季节性流感病毒正在全球范围内共同传播。一种能够快速、灵敏和特异性地区分大流行流感 A/H1N1 2009 病毒与季节性流感病毒，并对季节性流感 A 病毒进行分型的检测方法，将有助于这些病毒感染的监测和控制。本研究开发了一种用于鉴定 A 型和 B 型流感病毒以及大流行流感 A/H1N1 2009 的多重实时 RT-PCR（rRT-PCR）检测方法。该方法还可以同时对季节性流感 A 病毒进行亚型分型。分析灵敏度为 10-10(4)拷贝/反应。组间和组内变异系数分别为 0.04-0.45%和 0.08-0.97%。与传统 PCR 技术相比，新的多重 rRT-PCR 检测方法在季节性流感病毒的亚型分型方面更灵敏。本研究开发的 rRT-PCR 方法检测大流行流感 A/H1N1 2009 的结果与美国 CDC 的 rRT-PCR 检测协议高度一致（96.88%）。使用美国 CDC 的 rRT-PCR 检测方法被鉴定为“大流行流感 A/H1N1 2009 阳性”的样本，使用本方法正确地重新分类为 H3N2 亚型。综上所述，这种新的多重 rRT-PCR 方案可能成为改进大流行流感 A/H1N1 2009 和季节性流感病毒诊断和管理的重要工具。
