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

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  • DONG Yiyang
    Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology
  • XU Yan
    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)
  • LIU Zaixin
    Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory
  • FU Yuanfang
    Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory
  • OHASHI Toshinori
    Institute of Microchemical Technology (IMT) Co., Ltd.
  • MAWATARI Kazuma
    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
  • KITAMORI Takehiko
    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

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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.

Journal

  • Analytical Sciences

    Analytical Sciences 30 (3), 359-363, 2014

    The Japan Society for Analytical Chemistry

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