title: Determination of cattle foot-and-mouth disease virus by micro-ELISA method.
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.
authors.affiliation: Beijing Key Laboratory of Bioprocess, College of Life Science and Technology, Beijing University of Chemical Technology.
authors.forename: Yiyang
authors.initials: Y
authors.lastname: Dong
authors.forename: Yan
authors.initials: Y
authors.lastname: Xu
authors.forename: Zaixin
authors.initials: Z
authors.lastname: Liu
authors.forename: Yuanfang
authors.initials: Y
authors.lastname: Fu
authors.forename: Toshinori
authors.initials: T
authors.lastname: Ohashi
authors.forename: Kazuma
authors.initials: K
authors.lastname: Mawatari
authors.forename: Takehiko
authors.initials: T
authors.lastname: Kitamori
mesh_terms: D000818:Animals; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D005536:Foot-and-Mouth Disease / Q000175:diagnosis / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000302:isolation & purification*; D056656:Lab-On-A-Chip Devices