Determination of Cattle Foot-and-Mouth Disease Virus by Micro-ELISA Method

Yiyang DONG; Yan XU; Zaixin LIU; Yuanfang FU; Toshinori OHASHI; Kazuma MAWATARI; Takehiko KITAMORI

doi:10.2116/analsci.30.359

Original Papers

Determination of Cattle Foot-and-Mouth Disease Virus by Micro-ELISA Method

Yiyang DONG, Yan XU, Zaixin LIU, Yuanfang FU, Toshinori OHASHI, Kazuma MAWATARI, Takehiko KITAMORI

Author information

Yiyang DONG
Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology
Yan XU
Nanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka Prefecture University
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
Zaixin LIU
Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory
Yuanfang FU
Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory
Toshinori OHASHI
Institute of Microchemical Technology (IMT) Co., Ltd.
Kazuma MAWATARI
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
Department of Applied Chemistry, School of Engineering, The University of Tokyo
Takehiko KITAMORI
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST)
Department of Applied Chemistry, School of Engineering, The University of Tokyo

Keywords: Microchip, micro-ELISA, foot-and-mouth disease virus

JOURNAL FREE ACCESS

2014 Volume 30 Issue 3 Pages 359-363

DOI https://doi.org/10.2116/analsci.30.359

Details

Abstract

The development of foot-and-mouth disease virus (FMDV) detection methods is crucial for animal food security, tackling regional FMDV epidemic, and global FMDV prognostic control. For these purposes, a fast and sensitive analysis method is required. In this study, we developed a microchip-based ELISA (enzyme-linked immunosorbent assay), micro-ELISA, to realize FMDV detection. Nickel(II) chelating chemistry was utilized to immobilize recombinant protein (antigen) on polystyrene micro-beads in order to determine FMDV antibodies in cattle serum samples. In addition, reaction protocol and conditions were investigated. As a result, the FMDV detection was successfully demonstrated with only a 10-μL sample volume in 25-minute assay time. Analytical sensitivity was evaluated by a maximum nominal positiveness percentage value (NPPV) of 303 and a dilution factor of 32×. The method’s inter-run and intra-run CV (coefficients of variance) values were 15.5 and 17.1%, respectively, which were fully compatible with the OIE (World Organization for Animal Health) principle of validation of diagnosis assays for infectious diseases. The developed method should become a powerful tool for determining other animal contagious diseases and/or zoonosis.

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