title: Foot-and-mouth disease virus antigen detection enzyme-linked immunosorbent assay using multiserotype-reactive monoclonal antibodies.
abstract: Monoclonal antibody (MAb)-based sandwich direct enzyme-linked immunosorbent assay (MSD-ELISA) methods that can detect foot-and-mouth disease virus (FMDV) antigens, both multiserotype (MSD-ELISA/MS) (for O, A, C, and Asia 1) and single-serotype (MSD-ELISA/SS) (for O, A, and Asia 1, specifically), were developed. MAb 1H5 was used as an antigen-trapping antibody that reacted with all seven serotypes of FMDV. The MAbs 71F2, 70C4, 16C6, and 7C2 were used as peroxidase-labeled detecting antibodies for multiple serotypes (O, A, C, and Asia 1), type O, type A, and type Asia 1, respectively, in both MSD-ELISA/MS and MSD-ELISA/SS. Our MSD-ELISAs showed high specificity. They produced a very low background of negative samples (buffer, plasma, and saliva) and were able to detect FMDV antigens from clinical samples (plasma and saliva), with results correlating with those of real-time reverse transcription-PCR. In terms of sensitivity, the MSD-ELISAs showed higher optical density values against each diluted serotype antigen than the indirect sandwich ELISA method, which is currently recommended in the manual of the World Organization for Animal Health. The sensitivity and specificity of the MSD-ELISAs seem to be sufficient for the antigenic diagnosis of FMDV.
authors.affiliation: Exotic Diseases Research Station, National Institute of Animal Health, 6-20-1 Josuihoncho Kodaira, Tokyo 187-0022, Japan. morioka@affrc.go.jp
authors.forename: Kazuki
authors.initials: K
authors.lastname: Morioka
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Fukai
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yoshida
authors.forename: Reiko
authors.initials: R
authors.lastname: Yamazoe
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Onozato
authors.forename: Seiichi
authors.initials: S
authors.lastname: Ohashi
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Tsuda
authors.forename: Kenichi
authors.initials: K
authors.lastname: Sakamoto
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000302:isolation & purification; D000956:Antigens, Viral / Q000032:analysis* / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology / Q000302:isolation & purification*; D009195:Peroxidase; D010949:Plasma / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012463:Saliva / Q000821:virology; D012680:Sensitivity and Specificity; D012703:Serotyping; D013194:Staining and Labeling; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis / Q000821:virology
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D009195:Peroxidase