Mizuike Rika, Sasaki Tadahiro, Baba Koichi, Iwamoto Hisahiko, Shibai Yusuke, Kosaka Mieko, Kubota-Koketsu Ritsuko, Yang Cheng-Song, Du Anariwa, Sakudo Akikazu, Tsujikawa Muneo, Yunoki Mikihiro, Ikuta Kazuyoshi
Department of Virology, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Clin Vaccine Immunol. 2011 Mar;18(3):494-9. doi: 10.1128/CVI.00269-10. Epub 2011 Jan 12.
Since its emergence in April 2009, pandemic influenza A virus H1N1 (H1N1 pdm), a new type of influenza A virus with a triple-reassortant genome, has spread throughout the world. Initial attempts to diagnose the infection in patients using immunochromatography (IC) relied on test kits developed for seasonal influenza A and B viruses, many of which proved significantly less sensitive to H1N1 pdm. Here, we prepared monoclonal antibodies that react with H1N1 pdm but not seasonal influenza A (H1N1 and H3N2) or B viruses. Using two of these antibodies, one recognizing viral hemagglutinin (HA) and the other recognizing nucleoprotein (NP), we developed kits for the specific detection of H1N1 pdm and tested them using clinical specimens of nasal wash fluid or nasopharyngeal fluid from patients with influenza-like illnesses. The specificities of both IC test kits were very high (93% for the HA kit, 100% for the NP kit). The test sensitivities for detection of H1N1 pdm were 85.5% with the anti-NP antibody, 49.4% with the anti-HA antibody, and 79.5% with a commercially available influenza A virus detection assay. Use of the anti-NP antibody could allow the rapid and accurate diagnosis of H1N1 pdm infections.
自2009年4月出现以来，甲型H1N1流感大流行病毒（H1N1 pdm），一种具有三重重组基因组的新型甲型流感病毒，已在全球传播。最初尝试使用免疫层析法（IC）诊断患者感染时，依赖于为季节性甲型和乙型流感病毒开发的检测试剂盒，其中许多对H1N1 pdm的敏感性明显较低。在此，我们制备了与H1N1 pdm反应但不与季节性甲型流感（H1N1和H3N2）或乙型流感病毒反应的单克隆抗体。使用其中两种抗体，一种识别病毒血凝素（HA），另一种识别核蛋白（NP），我们开发了用于特异性检测H1N1 pdm的试剂盒，并使用流感样疾病患者的鼻洗液或鼻咽液临床标本对其进行了测试。两种IC检测试剂盒的特异性都非常高（HA试剂盒为93%，NP试剂盒为100%）。检测H1N1 pdm的检测灵敏度，抗NP抗体为85.5%，抗HA抗体为49.4%，市售甲型流感病毒检测试验为79.5%。使用抗NP抗体可实现对H1N1 pdm感染的快速准确诊断。
