title: Nanobodies, a promising tool for species-specific diagnosis of Taenia solium cysticercosis.
abstract: Taenia solium cysticercosis is a major helminth zoonosis in developing countries. Pigs are the intermediate hosts mediating transmission of infection. Specific assays to diagnose living cysts in pigs are lacking. The monoclonal-based antigen detection ELISA is genus-specific and cross-reactions with Taenia hydatigena hamper the use of this test to screen pigs. We, therefore, aimed to introduce nanobodies, camelid-derived single-domain antibodies specific for T. solium cysticercosis, to develop unambiguous tests. Nanobodies were cloned following immunization of two dromedaries with T. solium antigen and eight T. solium-specific nanobodies were selected after phage display. Their binding characteristics and potential for the diagnosis of porcine cysticercosis were investigated. The nanobodies do not cross-react with T. hydatigena, Taenia saginata, Taenia crassiceps or Trichinella spiralis and were categorized into four epitope-binding groups. The target protein was identified as 14kDa diagnostic glycoprotein (Ts14), but the nanobodies also reacted with other proteins of the same family. Nanobodies were tested in a sandwich ELISA with cyst fluid, and one particular nanobody detected its cognate serum antigens in a species-specific inhibition ELISA. Considering their beneficial production and stability properties, these highly specific nanobodies constitute a promising tool to diagnose cysticercosis after further improvement of the sensitivity and future assay validation.
authors.affiliation: Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium. ndeckers@itg.be
authors.forename: N
authors.initials: N
authors.lastname: Deckers
authors.forename: D
authors.initials: D
authors.lastname: Saerens
authors.forename: K
authors.initials: K
authors.lastname: Kanobana
authors.forename: K
authors.initials: K
authors.lastname: Conrath
authors.forename: B
authors.initials: B
authors.lastname: Victor
authors.forename: U
authors.initials: U
authors.lastname: Wernery
authors.forename: J
authors.initials: J
authors.lastname: Vercruysse
authors.forename: S
authors.initials: S
authors.lastname: Muyldermans
authors.forename: P
authors.initials: P
authors.lastname: Dorny
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000097:blood / Q000276:immunology; D015153:Blotting, Western / Q000379:methods; D002162:Camelus / Q000469:parasitology; D003429:Cross Reactions; D003551:Cysticercosis / Q000175:diagnosis / Q000469:parasitology / Q000662:veterinary*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D018604:Epitope Mapping / Q000379:methods; D005240:Feasibility Studies; D008969:Molecular Sequence Data; D053758:Nanoparticles; D012698:Serologic Tests / Q000379:methods; D013045:Species Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000469:parasitology; D041201:Taenia solium / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth