title: Generation of mAbs to foot-and-mouth disease virus serotype A and application in a competitive ELISA for serodiagnosis.
abstract: Foot-and-mouth disease (FMD) is an economically devastating disease that severely limits international trade of animals. Of the seven FMD virus (FMDV) serotypes, serotype A is one of the most widespread cross the world. Currently antibodies to FMDV are detected in animals using the virus neutralization test (VNT) and the enzyme-linked immunosorbent assay (ELISA). The VNT is laborious, time-consuming and reliant on live virus and cell cultures, while ELISA has the advantage of using inactivated antigens and often provides more reproducible results. The aim of this study was to develop a reliable and rapid competitive ELISA (cELISA) for the detection of antibodies to FMDV serotype A (FMDV/A).
  
  
  
  A panel of FMDV/A specific monoclonal antibodies (mAbs) was generated and their ability to compete with a polyclonal serum from FMDV/A-infected cattle was examined. Two mAbs inhibited the binding of a polyclonal serum to FMDV/A viruses. The binding epitopes of each were determined as conformational and located on the VP2 viral capsid protein. The FMDV/A cELISA was developed using these two mAbs and FMDV/A inactivated virus as antigen. The diagnostic specificity and sensitivity were 99.7 and 99.3% (98.5-100%) respectively, based on a predetermined cut-off of 50% inhibition. When analysing sera from animals experimentally infected with FMDV/A, the cELISA detected antibodies from 5-days post infection (dpi) and remained positive for at least 21-28 days post infection. Comparison based on the Kappa coefficient showed strong agreement (90-94%) between cELISA and VNT.
  
  
  
  The cELISA results are comparable to the VNT for antibody detection making it a simple and reliable test to detect antibodies against FMDV/A.
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada. Ming.Yang@inspection.gc.ca.
authors.forename: Ming
authors.initials: M
authors.lastname: Yang
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada.
authors.forename: Wanhong
authors.initials: W
authors.lastname: Xu
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada.
authors.forename: Hilary
authors.initials: H
authors.lastname: Bittner
authors.affiliation: Australian Animal Health Laboratory, CSIRO, 5 Portarlington Road, Geelong, VIC, 3220, Australia.
authors.forename: Jacquelyn
authors.initials: J
authors.lastname: Horsington
authors.affiliation: Australian Animal Health Laboratory, CSIRO, 5 Portarlington Road, Geelong, VIC, 3220, Australia.
authors.forename: Wilna
authors.initials: W
authors.lastname: Vosloo
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada.
authors.forename: Melissa
authors.initials: M
authors.lastname: Goolia
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada.
authors.forename: Diana
authors.initials: D
authors.lastname: Lusansky
authors.affiliation: National Centre for Foreign Animal Disease, 1015 Arlington Street, Winnipeg, MB, R3E 3M4, Canada.
authors.forename: Charles
authors.initials: C
authors.lastname: Nfon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000914:Antibodies, Viral / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005536:Foot-and-Mouth Disease / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology*; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral
keywords: Competitive ELISA; Foot–and–mouth disease virus; Monoclonal antibody; Serodiagnosis; Serotype A