title: Cloning and characterization of the Leishmania (Viannia) braziliensis Hsp70 gene. Diagnostic use of the C-terminal fragment rLb70(513-663).
abstract: Cross-reactions between Leishmania braziliensis and Trypanosoma cruzi caused by common antigenic determinants hinder the specific diagnosis of cutaneous and mucocutaneous leishmaniasis (CL and MCL). Therefore, the usefulness of the 70-kDa heat shock protein (Hsp70) from L. braziliensis for differential serodiagnosis was investigated. The single-copy gene encoding Hsp70, consisting of 663 amino acids, was isolated from a genomic DNA library. The antigenicity data show that Hsp70 is an immunodominant antigen highly recognized (84%) by sera of patients with CL and MCL and to a lesser extent by chagasic patients (18.75%). Antigenic mapping of the 5 overlapping fragments into which the protein was split showed that the main antigenic determinants are located in the carboxy-terminal end. The linear antigenic determinants that show cross-reactions with chagasic sera are located in the fragment rLb70(352-518). The carboxy-terminal fragment rLb70(513-663) presents 70% sensitivity and 100% specificity, so it could be a potential candidate for specific serodiagnosis of CL and MCL caused by L. braziliensis.
authors.affiliation: Departamento de Parasitología, Facultad de Farmacia, Universidad de La Laguna, c/ Astrofísico Francisco Sánchez s/n, Tenerife 38271, Spain.
authors.forename: Ada I
authors.initials: AI
authors.lastname: Zurita
authors.forename: Javier
authors.initials: J
authors.lastname: Rodríguez
authors.forename: José E
authors.initials: JE
authors.lastname: Piñero
authors.forename: Rosa
authors.initials: R
authors.lastname: Pacheco
authors.forename: Emma
authors.initials: E
authors.lastname: Carmelo
authors.forename: Antonio
authors.initials: A
authors.lastname: del Castllo
authors.forename: Basilio
authors.initials: B
authors.lastname: Valladares
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology*; D015139:Blotting, Southern; D014355:Chagas Disease / Q000175:diagnosis; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003429:Cross Reactions; D003937:Diagnosis, Differential; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007892:Leishmania braziliensis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis*; D007897:Leishmaniasis, Mucocutaneous / Q000175:diagnosis*; D008969:Molecular Sequence Data; D016366:Open Reading Frames / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
chemical_list: D000953:Antigens, Protozoan; D000939:Epitopes; D018840:HSP70 Heat-Shock Proteins; D007106:Immune Sera; D011994:Recombinant Proteins