title: Expression of a recombinant protein, A2 family, from Leishmania infantum (Jaboticabal strain) and its evaluation in Canine Visceral Leishmaniasis serological test.
abstract: This study aimed to express a recombinant A2 family protein of Leishmania chagasi, Jaboticabal strain; test this protein as an antigen in serological assays; and investigate its antigenicity and immunogenicity. A protein coded by an allele of the A2 gene isolated from L. chagasi was expressed in three different strains of Escherichia coli. We used 29 sera samples from Leishmune-vaccinated dogs, 482 sera samples from dogs from endemic areas (positive controls), and 170 sera samples from dogs from non-endemic areas (negative controls) in ELISA tests using soluble Leishmaniaantigen (SLA) and His-A2 as antigen. Expressed proteins showed, by western blotting, the expression of an 11 KDa protein. Sixty-three percent (303/482) of the samples from endemic areas were positive by ELISA His-A2, whereas 93.1% (27/29) of Leishmune®-vaccinated animals were negative by His-A2-ELISA. Anti-A2 antibodies from mice inoculated with the A2 protein were detected in slides containing amastigote forms, but not in slides containing promastigote forms. The A2 recombinant protein from L. chagasi may be a useful tool in the diagnosis of CVL, and further tests regarding the infection stage and the specie of parasite at which the dogs are sampled should provide a better understanding of our results.
authors.affiliation: Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP, BR.
authors.forename: Márcia Mariza Gomes
authors.initials: MM
authors.lastname: Jusi
authors.affiliation: Departamento de Zootecnia, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, BR.
authors.forename: Trícia Maria Ferreira de Sousa
authors.initials: TM
authors.lastname: Oliveira
authors.affiliation: Faculdade de Ciências Agrárias e Sociais, Itapeva, SP, BR.
authors.forename: Andréa Cristina Higa
authors.initials: AC
authors.lastname: Nakaghi
authors.affiliation: Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP, BR.
authors.forename: Marcos Rogério
authors.initials: MR
authors.lastname: André
authors.affiliation: Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal, SP, BR.
authors.forename: Rosangela Zacarias
authors.initials: RZ
authors.lastname: Machado
mesh_terms: D000818:Animals; D000953:Antigens, Protozoan / Q000097:blood; D004283:Dog Diseases / Q000097:blood* / Q000175:diagnosis*; D004285:Dogs; D018314:Leishmania infantum / Q000276:immunology / Q000378:metabolism*; D007898:Leishmaniasis, Visceral / Q000097:blood / Q000662:veterinary*; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000097:blood*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000097:blood; D012698:Serologic Tests
chemical_list: D000953:Antigens, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins