title: A promising new ELISA diagnostic test for cattle babesiosis based on Babesia bigemina Apical Membrane Antigen-1.
abstract: Babesiosis due to Babesia bigemina is a relevant tick-borne disease, affecting cattle worldwide. Many surface proteins of the pathogen including the Apical Membrane Antigen 1 (AMA-1) - have been analysed for vaccine and diagnostic purposes. This study focused on B. bigemina AMA-1 and on its use for the assessment of diagnostic tests. After bioinformatic analyses, AMA-1 codifying region was amplified and cloned into an expression vector used to induce protein synthesis in Escherichia coli cells. AMA-1 was purified by affinity chromatography and used to set up the best condition for an ELISA protocol. Bovine field sera positive to B. bigemina were used to evaluate the presence of anti-AMA-1 antibodies. In order to verify the assay specificity, sera positive to Babesia bovis or to the piroplasm Theileria annulata were also included. Significant differences were obtained between sera negative to both B. bigemina and B. bovis and samples positive to B. bigemina, to B. bovis or to both pathogens. No significant reaction was observed with T. annulata positive sera. The results showed that AMA-1 protein is suitable to be used as antigen in diagnostic assays for babesiosis diagnosis in cattle, as it does not show any cross reaction with anti-T. annulata antibodies.
authors.affiliation: Istituto Zoopro lattico Sperimentale della Sicilia, Via G. Marinuzzi 3, 90129 Palermo, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Torina
authors.forename: Antonio
authors.initials: A
authors.lastname: Cordaro
authors.forename: Valeria
authors.initials: V
authors.lastname: Blanda
authors.forename: Rosalia
authors.initials: R
authors.lastname: D'Agostino
authors.forename: Salvatore
authors.initials: S
authors.lastname: Scimeca
authors.forename: Maria E
authors.initials: ME
authors.lastname: Scariano
authors.forename: Guido
authors.initials: G
authors.lastname: Sireci
authors.forename: Rossella
authors.initials: R
authors.lastname: Lelli
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis*; D001403:Babesia / Q000276:immunology*; D001404:Babesiosis / Q000175:diagnosis*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D008565:Membrane Proteins / Q000032:analysis*; D015800:Protozoan Proteins / Q000032:analysis*
chemical_list: D000953:Antigens, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins