Rapid detection of anti-H5 avian influenza virus antibody by fluorescence polarization immunoassay using a portable fluorescence polarization analyzer

Keine Nishiyama; Yohei Takeda; Masatoshi Maeki; Akihiko Ishida; Hirofumi Tani; Koji Shigemura; Akihide Hibara; Yutaka Yonezawa; Kunitoshi Imai; Haruko Ogawa; Manabu Tokeshi

doi:10.1016/j.snb.2020.128160

Rapid detection of anti-H5 avian influenza virus antibody by fluorescence polarization immunoassay using a portable fluorescence polarization analyzer

Sens Actuators B Chem. 2020 Aug 1:316:128160. doi: 10.1016/j.snb.2020.128160. Epub 2020 Apr 21.

Authors

Affiliations

¹ Graduate School of Chemical Sciences and Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan.
² Research Center for Global Agromedicine, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada, Obihiro, Hokkaido 080-8555, Japan.
³ Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan.
⁴ Tianma Japan, Ltd., Shin-Kawasaki Mitsui Building West Tower 28F 1-1-2, Kashimada, Saiwai-ku, Kawasaki, Kanagawa 212-0058, Japan.
⁵ Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai 980-8577, Japan.
⁶ Department of Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine, 2-11 Inada, Obihiro, Hokkaido 080-8555, Japan.

Abstract

A rapid, facile and selective detection of anti-H5 subtype avian influenza virus (AIV) antibody in serum by fluorescence polarization immunoassay (FPIA) was achieved. A fragment of recombinant H5 subtype AIV hemagglutinin was produced and labeled with fluorescein to use it as a labeled antigen in FPIA. This labeled antigen was mixed with anti-AIV sera (H1-H16 subtypes) and FP of the mixture was measured using a portable FP analyzer on a microdevice. It was found that FP increased in proportion to the concentration of anti-H5 AIV antibody (serum) and was significantly higher than FP obtained with the other sera. The selective detection of anti-H5 subtype AIV antibody was confirmed. The required volume of original sample was 2 μL and analysis time was within 20 min. This detection system realizes an efficient on-site diagnosis and surveillance of AIV.

Keywords: Antibody detection; Avian influenza; Fluorescence polarization immunoassay; Rapid diagnosis.