title: Development of a multiplex lateral flow strip test for foot-and-mouth disease virus detection using monoclonal antibodies.
abstract: Foot-and-mouth disease (FMD) is one of the world's most highly contagious animal diseases with tremendous economic consequences. A rapid and specific test for FMD diagnosis at the site of a suspected outbreak is crucial for the implementation of control measures. This project developed a multiplex lateral flow immunochromatographic strip test (multiplex-LFI) for the rapid detection and serotyping of FMD viruses. The monoclonal antibodies (mAbs) against serotypes O, A, and Asia 1 were used as capture mAbs. The mAbs were conjugated with fluorescein, rhodamine or biotin for serotype O, A and Asia 1, respectively. The detection mAbs which consisted of a serotype-independent mAb in combination with one serotype A-specific mAb and one Asia 1-specific mAb, were each colloidal gold-conjugated. The strips used in this study contained one control line and three test lines, which corresponded to one of the three serotypes, O, A or Asia 1. The newly developed multiplex-LFI strip test specifically identified serotype O (n=46), A (n=45) and Asia 1 (n=17) in all tested field isolates. The sensitivity of this strip test was comparable to the double antibody sandwich ELISA for serotypes O and A, but lower than the ELISA for serotype Asia 1. The multiplex-LFI strip test identified all tissue suspensions from animals that were experimentally inoculated with serotypes O, A or Asia 1. FMD viruses were detected in 38% and 50% of the swab samples from the lesion areas of experimentally inoculated sheep for serotypes O and A, respectively. The capability of the multiplex-LFI strip tests to produce rapid results with high specificity for FMD viruses of multiple serotypes makes this test a valuable tool to detect FMD viruses at outbreak sites.
authors.affiliation: National Centre for Foreign Animal Disease, Canadian food Inspection Agency, Winnipeg, Manitoba, Canada. Electronic address: Ming.Yang@inspection.gc.ca.
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.affiliation: RapidAssays ApS, Gunloegsgade, Copenhagen-S, Denmark. Electronic address: rapidassays@gmail.com.
authors.forename: Nigel R
authors.initials: NR
authors.lastname: Caterer
authors.affiliation: National Centre for Foreign Animal Disease, Canadian food Inspection Agency, Winnipeg, Manitoba, Canada. Electronic address: Wanhong.Xu@inspection.gc.ca.
authors.forename: Wanhong
authors.initials: W
authors.lastname: Xu
authors.affiliation: National Centre for Foreign Animal Disease, Canadian food Inspection Agency, Winnipeg, Manitoba, Canada. Electronic address: Melissa.Goolia@inspection.gc.ca.
authors.forename: Melissa
authors.initials: M
authors.lastname: Goolia
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology; D001208:Asia; D002846:Chromatography, Affinity / Q000379:methods*; D005536:Foot-and-Mouth Disease / Q000175:diagnosis*; D029544:Foot-and-Mouth Disease Virus / Q000145:classification* / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods*; D014730:Veterinary Medicine / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral
keywords: Foot-and-mouth disease; Monoclonal antibodies; Multiplex lateral flow immunochromatographic strip test; Rapid virus detection