title: Development of a competitive ELISA for NS3 antibodies as DIVA test accompanying the novel Disabled Infectious Single Animal (DISA) vaccine for Bluetongue.
abstract: Recently, we have developed a novel vaccine for Bluetongue named BT Disabled Infectious Single Animal (DISA) vaccine. Due to the lack of non-essential NS3/NS3a protein, BT DISA vaccine is a replicating vaccine, but without the inherent risks of live-attenuated vaccines, such as residual virulence or reversion to virulence by mutations, reassortment with field virus, horizontal spread by vectors and vertical transmission. The immune response induced by BT DISA vaccines is rapidly induced, highly protective and serotype specific which is dependent on the immunodominant and serotype determining VP2 protein. The BT DISA vaccine platform provides the replacement of exclusively VP2 from different serotypes in order to safely formulate multivalent cocktail vaccines. The lack of NS3/NS3a directed antibodies by BT DISA vaccination enables differentiation of infected from vaccinated animals (DIVA principle). A highly conserved immunogenic site corresponding to the late domain was mapped in the N-terminal region of NS3. We here established an NS3-specific competitive ELISA (NS3 cELISA) as serological DIVA test accompanying BT DISA vaccines. To this end, NS3 protein missing putative transmembrane regions was produced in large amounts in bacteria and used as antigen in the NS3 cELISA which was investigated with a variety of sera. The NS3 cELISA displayed a high sensitivity and specificity similar to the commercially available VP7-specific cELISA. Results of previously performed vaccination-challenge trials with BT DISA vaccines clearly demonstrate the DIVA system based on the NS3 cELISA and BT vaccine free of NS3 protein.
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
authors.forename: Mirriam G J
authors.initials: MGJ
authors.lastname: Tacken
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
authors.forename: Franz J
authors.initials: FJ
authors.lastname: Daus
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands; Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
authors.forename: Femke
authors.initials: F
authors.lastname: Feenstra
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
authors.forename: René G P
authors.initials: RGP
authors.lastname: van Gennip
authors.affiliation: Department of Virology, Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands; Department of Biochemistry, Centre for Human Metabonomics, North-West University, Potchefstroom, South Africa. Electronic address: piet.vanrijn@wur.nl.
authors.forename: Piet A
authors.initials: PA
authors.lastname: van Rijn
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood*; D001820:Bluetongue virus; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D008969:Molecular Sequence Data; D012680:Sensitivity and Specificity; D012756:Sheep; D017361:Viral Nonstructural Proteins / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; C075080:S10 protein, Bluetongue virus; D017361:Viral Nonstructural Proteins; D014765:Viral Vaccines
keywords: Bluetongue; DIVA; Disabled Infectious Single Animal vaccine; ELISA; Non-structural protein 3