Rapid, Sensitive, and Selective Detection of H5 Hemagglutinin from Avian Influenza Virus Using an Immunowall Device

Kenia Chávez Ramos; Keine Nishiyama; Masatoshi Maeki; Akihiko Ishida; Hirofumi Tani; Toshihiro Kasama; Yoshinobu Baba; Manabu Tokeshi

doi:10.1021/acsomega.9b02788

Rapid, Sensitive, and Selective Detection of H5 Hemagglutinin from Avian Influenza Virus Using an Immunowall Device

ACS Omega. 2019 Sep 27;4(15):16683-16688. doi: 10.1021/acsomega.9b02788. eCollection 2019 Oct 8.

Authors

Kenia Chávez Ramos¹, Keine Nishiyama², Masatoshi Maeki², Akihiko Ishida², Hirofumi Tani², Toshihiro Kasama^{3

4}, Yoshinobu Baba^{4

5}, Manabu Tokeshi^{2

4}

Affiliations

¹ Department of Analytical Chemistry, School of Chemistry, National Autonomous University of Mexico, Av. Universidad 3000, Mexico City 04510, Mexico.
² Graduate School of Chemical Sciences and Engineering, and Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, Kita 13 Nishi 8, Kita-ku, Sapporo 060-8628, Japan.
³ Department of Bioengineering, School of Engineering, The University of Tokyo, Shinkawasaki, Saiwai-ku, Kanagawa 212-0032, Japan.
⁴ Institute of Nano-Life Systems, Institutes of Innovation for Future Society, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8603, Japan.
⁵ Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya 464-8603, Japan.

Abstract

Avian influenza virus (AIV) infection, caused by influenza virus type A, is an infectious, acute respiratory disease of birds related to influenza outbreaks worldwide. The highly pathogenic AIV subtype H5N1 has crossed species barriers to infect mammals, including humans, with fatal outcomes and has received attention as a potential pandemic threat. A rapid and timely detection in poultry is vitally important to prevent the virus spread. Despite their great sensitivity, conventional detection methods such as real-time reverse transcription-polymerase chain reaction and the agar gel precipitation test are time-consuming and labor-intensive and require special training. In this work, an immunowall device was evaluated as an easier and faster way for detecting AIV H5-hemagglutinin (AIV H5-HA). For detection, fluorescence-labeled or enzyme-labeled antibody was employed as a labeling antibody in a sandwich immunoassay. Both were shown in this paper to be easier and faster assays for detection compared with the conventional enzyme-linked immunosorbent assay (ELISA) kit. In addition, high selectivity was achieved for AIV H5-HA detection after the evaluation of other different HA virus subtypes. The limit of detection was 0.23 ng/mL for the enzyme-labeled antibody. This value was equivalent to that of the conventional ELISA kit but 8 times faster (31 min compared to 260 min). The detection range was 0.23-100 ng/mL. The immunowall device with the enzyme-labeled antibody offers a rapid, sensitive, selective, and simple immunoassay system for future H5 AIV real sample detection.