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

Keine Nishiyama, Yohei Takeda, Kazuki Takahashi, Mao Fukuyama, Masatoshi Maeki, Akihiko Ishida, Hirofumi Tani, Koji Shigemura, Akihide Hibara, Haruko Ogawa, Manabu Tokeshi

Research output: Contribution to journalArticlepeer-review

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 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.

Original languageEnglish
JournalAnalytical and Bioanalytical Chemistry
DOIs
Publication statusAccepted/In press - 2021

Keywords

  • Avian influenza
  • Fluorescence polarization immunoassay
  • Rapid diagnosis
  • Virus detection

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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