title: Microplate Agglutination Test for Canine Brucellosis Using Recombinant Antigen-Coated Beads.
abstract: Brucella canis, a facultative intracellular pathogen, is the causative agent of canine brucellosis. The diagnosis of canine brucellosis is based on bacteriological examination and serological methods, including agglutination and gel diffusion tests. In this study, four recombinant antigens, heat shock protein 60, rhizopine-binding protein, Cu-Zn superoxide dismutase, and hypothetical protein (Ag 4), were constructed. These antigens were coated on latex beads and their usefulness in the serological diagnosis of canine brucellosis was examined. All recombinant antigens showed specific reaction with sera from B. canis-infected dogs in Western blotting. In a microplate agglutination test, mixing sera from B. canis-infected dogs, but not sera from B. canis-free dogs, with single or multiple antigens-coated latex beads produced clear agglutination. Moreover, the antigen-coated latex beads did not show nonspecific agglutination in hemolyzed serum samples. A survey of canine serum samples conducted by the microplate agglutination test using single antigen-coated latex beads showed that this method would be useful in the serological diagnosis of canine brucellosis. Further investigations using more serum samples are required to confirm the usefulness of our method.
authors.affiliation: The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Yussaira
authors.identifier: 0000-0003-3512-2676
authors.initials: Y
authors.lastname: Castillo
authors.affiliation: The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Masato
authors.initials: M
authors.lastname: Tachibana
authors.affiliation: Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Yui
authors.initials: Y
authors.lastname: Kimura
authors.affiliation: Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Suk
authors.initials: S
authors.lastname: Kim
authors.affiliation: Merial Japan Limited, 3-20-2 Nishi Shinjuku, Shinjuku, Tokyo 163-1488, Japan.
authors.forename: Yasuaki
authors.initials: Y
authors.lastname: Ichikawa
authors.affiliation: Laboratory of Small Animal Internal Medicine, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Endo
authors.affiliation: The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan; Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Kenta
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan; Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Takashi
authors.identifier: 0000-0003-1294-0799
authors.initials: T
authors.lastname: Shimizu
authors.affiliation: The United Graduate School of Veterinary Science, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan; Laboratory of Veterinary Public Health, Joint Faculty of Veterinary Medicine, 1677-1 Yoshida, Yamaguchi 753-8515, Japan.
authors.forename: Masahisa
authors.identifier: 0000-0003-3477-3135
authors.initials: M
authors.lastname: Watarai