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Analyst . Detection of H5 subtype avian influenza virus in avian oropharyngeal swab samples using a microfluidic non-competitive fluorescence polari

tetano

Editor, Senior Moderator
Analyst


. 2026 Jun 17.
doi: 10.1039/d6an00505e. Online ahead of print.
Detection of H5 subtype avian influenza virus in avian oropharyngeal swab samples using a microfluidic non-competitive fluorescence polarization immunoassay

Kazuki Takahashi[SUP] 1 [/SUP], Yohei Takeda[SUP] 2 [/SUP], Keine Nishiyama[SUP] 1 [/SUP], Mao Fukuyama[SUP] 3 [/SUP], Masatoshi Maeki[SUP] 4 [/SUP], Akihiko Ishida[SUP] 4 [/SUP], Hirofumi Tani[SUP] 4 [/SUP], Ayuko Imai[SUP] 5 [/SUP], Koji Shigemura[SUP] 5 [/SUP], Akihide Hibara[SUP] 6 [/SUP], Haruko Ogawa[SUP] 2 [/SUP], Manabu Tokeshi[SUP] 4 7 8 [/SUP]


Affiliations
Abstract

Rapid and simple methods for on-site detection of avian influenza virus (AIV) are essential for effective field surveillance. In this study, a microfluidic device-based non-competitive fluorescence polarization immunoassay (NC-FPIA) using an ATTO 647N-labeled hemagglutinin fragment of H5 subtype AIV (H5-AIV) as a tracer was developed for the quantification of H5-AIV in avian oropharyngeal swab samples. Autofluorescence from the swab matrix significantly interfered with fluorescence polarization measurements. However, quantitative detection of H5-AIV was achieved by suppressing the matrix effect through 80-fold dilution. The limit of detection was 186.4 μg mL[SUP]-1[/SUP]. The assay can be completed within 20 minutes without complex sample preparation, making it suitable for rapid on-site applications, including field surveillance of AIV in wild birds and poultry. Furthermore, by designing appropriate tracers, the method can be extended to other AIV subtypes.


 
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