Graduate School of Chemical Sciences and Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo, 060-8628, Japan.
Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada, Obihiro, Hokkaido, 080-8555, Japan.
Anal Bioanal Chem. 2021 Jul;413(18):4619-4623. doi: 10.1007/s00216-021-03193-y. Epub 2021 Feb 5.
Nowadays, the diagnosis of viral infections is receiving broad attention. We have developed a non-competitive fluorescence polarization immunoassay (NC-FPIA), which is a separation-free immunoassay, for a virus detection. H5 subtype avian influenza virus (H5-AIV) was used as a model virus for the proof of concept. The fluorescein-labeled Fab fragment that binds to H5 hemagglutinin was used for NC-FPIA. The purified H5-AIV which has H5 hemagglutinin was mixed with the fluorescein-labeled Fab fragment. After that, the degree of fluorescence polarization was measured with a portable FPIA analyzer. H5-AIV was successfully detected with an incubation time of 15 min. In addition, the portable FPIA analyzer enables performance of on-site NC-FPIA with a sample volume of 20 μL or less. This is the first research of detecting a virus particle by FPIA. This NC-FPIA can be applied to rapid on-site diagnosis of various viruses.
如今，病毒感染的诊断受到广泛关注。我们开发了一种无分离的荧光偏振免疫分析（NC-FPIA），用于病毒检测。我们选择 H5 亚型禽流感病毒（H5-AIV）作为概念验证的模型病毒。用于 NC-FPIA 的是与 H5 血凝素结合的荧光素标记 Fab 片段。将具有 H5 血凝素的纯化 H5-AIV 与荧光素标记的 Fab 片段混合。之后，使用便携式 FPIA 分析仪测量荧光偏振程度。H5-AIV 在孵育 15 分钟后成功被检测到。此外，便携式 FPIA 分析仪可通过 20μL 或更少的样品量在现场进行 NC-FPIA 分析。这是首次通过 FPIA 检测病毒颗粒。这种 NC-FPIA 可应用于各种病毒的快速现场诊断。
