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Non-competitive fluorescence polarization immunoassay for detection of H5 avian influenza virus using a portable analyzer.


ABSTRACT: 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 ?L 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.

SUBMITTER: Nishiyama K 

PROVIDER: S-EPMC7864621 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Non-competitive fluorescence polarization immunoassay for detection of H5 avian influenza virus using a portable analyzer.

Nishiyama Keine K   Takeda Yohei Y   Takahashi Kazuki K   Fukuyama Mao M   Maeki Masatoshi M   Ishida Akihiko A   Tani Hirofumi H   Shigemura Koji K   Hibara Akihide A   Ogawa Haruko H   Tokeshi Manabu M  

Analytical and bioanalytical chemistry 20210205 18


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 frag  ...[more]

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