Sakudo Akikazu, Baba Koichi, Tsukamoto Megumi, Sugimoto Atsuko, Okada Takashi, Kobayashi Takanori, Kawashita Norihito, Takagi Tatsuya, Ikuta Kazuyoshi
Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Bioorg Med Chem. 2009 Jan 15;17(2):752-7. doi: 10.1016/j.bmc.2008.11.046. Epub 2008 Nov 24.
An anionic magnetic beads-based method was developed for the capture of human influenza A and B viruses from nasal aspirates, allantoic fluid and culture medium. A polymer, poly(methyl vinyl ether-maleic anhydride) [poly(MVE-MA)], was used to endow magnetic beads with a negative charge and bioadhesive properties. After incubation with samples containing human influenza virus, the beads were separated from supernatants by applying a magnetic field. The adsorption [corrected] of the virus by the beads was confirmed by hemagglutinin assay, immunochromatography, Western blotting, egg infection, and cell infection. Successful capture was proved using 5 H1N1 influenza A viruses, 10 H3N2 influenza A viruses, and 6 influenza B viruses. Furthermore, the infectivity in chicken embryonated eggs and Madin-Darby canine kidney (MDCK) cells of the captured human influenza virus was similar to that of the total viral quantity of starting materials. Therefore, this method of capture using magnetic beads coated with poly(MVE-MA) can be broadly used for the recovery of infectious human influenza viruses.
开发了一种基于阴离子磁珠的方法，用于从鼻洗液、尿囊液和培养基中捕获甲型和乙型人流感病毒。使用一种聚合物聚（甲基乙烯基醚-马来酸酐）[聚（MVE-MA）]赋予磁珠负电荷和生物粘附特性。在与含有人流感病毒的样品孵育后，通过施加磁场将磁珠与上清液分离。通过血凝素测定、免疫色谱法、蛋白质印迹法、鸡胚感染和细胞感染证实了磁珠对病毒的吸附。使用5株甲型H1N1流感病毒、10株甲型H3N2流感病毒和6株乙型流感病毒证明了捕获成功。此外，捕获的人流感病毒在鸡胚和Madin-Darby犬肾（MDCK）细胞中的感染性与起始材料的总病毒量相似。因此，这种使用涂有聚（MVE-MA）的磁珠的捕获方法可广泛用于回收具有传染性的人流感病毒。
