Analytical Sciences

Abstract − Analytical Sciences, 30(3), 359 (2014).

Determination of Cattle Foot-and-Mouth Disease Virus by Micro-ELISA Method
Yiyang DONG,*1 Yan XU,*2,*3 Zaixin LIU,*4 Yuanfang FU,*4 Toshinori OHASHI,*5 Kazuma MAWATARI,*3,*6 and Takehiko KITAMORI*3,*6
*1 Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology, 15 Beisanhuan East Road, Chaoyang District, Beijing 100029, P. R. China
*2 Nanoscience and Nanotechnology Research Center, Research Organization for the 21st Century, Osaka Prefecture University, 1-2 Gakuen, Naka, Sakai, Osaka 599-8570, Japan
*3 Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), 5 Sanban, Chiyoda, Tokyo 102-0075, Japan
*4 Lanzhou Veterinary Research Institute, Chinese Academy of Agriculture Science, State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Xujiaping No. 1, Yanchangpu, Lanzhou, Gansu 730046, P. R. China
*5 Institute of Microchemical Technology (IMT) Co., Ltd., 3-2-1 Sakado, Takatsu, Kawasaki, Kanagawa 213-0012, Japan
*6 Department of Applied Chemistry, School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo 113-8656, Japan
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.