title: Truncated Bovine Integrin Alpha-v/Beta-6 as a Universal Capture Ligand for FMD Diagnosis.
abstract: Foot-and-mouth disease (FMD) is endemic in many regions of the world and is one of the most prevalent epizootic animal diseases. FMD affects livestock, such as cattle, sheep, goats and pigs, and causes enormous economic losses due to reduced productivity and trade restrictions. Preparedness and early diagnosis are essential for effective control of FMD. Many diagnostic assays are dependent on raising high-affinity, anti-FMD virus (FMDV) serotype-specific antibodies in small animals (rabbits and guinea pigs) that give broad virus coverage. Here we show that soluble, truncated forms of bovine αvβ6 bind FMDV in an authentic RGD and divalent cation dependent interaction and can be used as the trapping reagent in a FMDV sandwich ELISA. In addition, inclusion of FLAG or His tags facilitates simple purification without the loss of virus binding. We also provide evidence that when combined with a guinea pig polyclonal serum, or serotype-specific monoclonal antibodies, the integrin can be used to detect viruses representative of all FMDV serotypes. We also show that recombinant FMDV empty capsids, with stabilising disulphide bonds, can serve as an antigen in the ELISA and can therefore replace inactivated virus antigen as a positive control for the assay. Our results demonstrate the potential use of bovine αvβ6 and FMDV empty capsids in FMD diagnostic assays.
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Gareth
authors: G
authors: Shimmon
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Britta A
authors: BA
authors: Wood
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Alison
authors: A
authors: Morris
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Valerie
authors: V
authors: Mioulet
authors: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Via Bianchi 9, Brescia, Italy.
authors: Santina
authors: S
authors: Grazioli
authors: Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Via Bianchi 9, Brescia, Italy.
authors: Emiliana
authors: E
authors: Brocchi
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Stephen
authors: S
authors: Berryman
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Toby
authors: T
authors: Tuthill
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Donald P
authors: DP
authors: King
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Alison
authors: A
authors: Burman
authors: The Pirbright Institute, Ash Rd, Pirbright, Surrey, GU24 0NF, United Kingdom.
authors: Terry
authors: T
authors: Jackson
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000914:Antibodies, Viral / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D002213:Capsid / Q000276:immunology*; D002417:Cattle; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000276:immunology / Q000302:isolation & purification*; D016023:Integrins / Q000276:immunology*; D011817:Rabbits
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000951:Antigens, Neoplasm; D016023:Integrins; C073878:integrin alphavbeta6