title: Comparative evaluation of enzyme-linked immunosorbent assays based on crude and recombinant leishmanial antigens for serodiagnosis of symptomatic and asymptomatic Leishmania infantum visceral infections in dogs.
abstract: The diagnosis of visceral leishmaniasis remains difficult in rural areas where the disease is endemic, and serologic methods still need assessment, as they are not very sensitive for the detection of asymptomatic infectious dogs. Here we present data on the development of enzyme-linked immunosorbent assay (ELISA)-based methods for the detection of antibodies against recombinant leishmanial antigens (namely, the recombinant K26 [rK26] and rK39 antigens from Leishmania infantum and the rA2 protein from Leishmania donovani) in comparison to ELISAs employing crude soluble antigen (CSA). The assays utilized sera from known negative controls (n=25) and clinically asymptomatic (n=50) and symptomatic (n=50) dogs with confirmed L. infantum infections. Additional studies were also done using sera from animals harboring other infections (n=14) for the evaluation of cross-reactivity. Our study indicated that rK26 and rK39 used in ELISAs provided very high sensitivities for the detection of symptomatic dogs (94% and 100%, respectively), followed by CSA (88%) and rA2 (70%). Conversely, rA2 was more sensitive for asymptomatic dogs (88%) than rK39 and rK26 (both 66%) and CSA (30%). Some cross-reactivity in sera from dogs with other infections (Leishmania braziliensis and Leptospira interrogans) was identified, but the rA2 protein provided the greatest specificity (98%). Data further indicate that all three recombinant proteins must be used in parallel to detect essentially all infected dogs. Efforts should be made to develop a cheap and reliable serologic test based on epitope selection from these diagnostic markers for the sensitive detection of L. infantum-infected dogs.
authors.affiliation: Department of Immunology, Oswaldo Cruz Institute, FIOCRUZ, Pav. Leonidas Deane, 5th Floor, Room 509, Av. Brasil 4365, CEP 21045-090 Rio de Janeiro, RJ, Brazil.
authors.forename: Renato
authors.initials: R
authors.lastname: Porrozzi
authors.forename: Marcos V
authors.initials: MV
authors.lastname: Santos da Costa
authors.forename: Antonio
authors.initials: A
authors.lastname: Teva
authors.forename: Aloísio
authors.initials: A
authors.lastname: Falqueto
authors.forename: Adelson L
authors.initials: AL
authors.lastname: Ferreira
authors.forename: Claudiney D
authors.initials: CD
authors.lastname: dos Santos
authors.forename: Ana Paula
authors.initials: AP
authors.lastname: Fernandes
authors.forename: Ricardo T
authors.initials: RT
authors.lastname: Gazzinelli
authors.forename: Antonio
authors.initials: A
authors.lastname: Campos-Neto
authors.forename: Gabriel
authors.initials: G
authors.lastname: Grimaldi
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood* / Q000276:immunology; D000953:Antigens, Protozoan; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007893:Leishmania donovani / Q000276:immunology*; D018314:Leishmania infantum / Q000276:immunology*; D007898:Leishmaniasis, Visceral / Q000175:diagnosis / Q000276:immunology* / Q000662:veterinary; D015800:Protozoan Proteins; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C107645:A2 protein, Leishmania donovani; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; C400962:K26 antigen, Leishmania chagasi; D015800:Protozoan Proteins; C079253:K39 antigen, Leishmania