Nanobodies, a promising tool for species-specific diagnosis of Taenia solium cysticercosis.
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.
Deckers
N
N
Animal Health Department, Institute of Tropical Medicine, Nationalestraat 155, B-2000 Antwerp, Belgium. ndeckers@itg.be
Saerens
D
D
Kanobana
K
K
Conrath
K
K
Victor
B
B
Wernery
U
U
Vercruysse
J
J
Muyldermans
S
S
Dorny
P
P
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*
D000909:Antibodies, Helminth; D000947:Antigens, Helminth