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BACKGROUND   Dogs are the main peridomiciliary reservoir of  Leishmania infantum  thus the correct diagnosis of infection is essential for the control of the transmission and treatment as well. However, the diagnosis is based on serological assays that are not fully effective.    OBJECTIVE   We aimed to establish an effective serological assay for the diagnosis of  L. infantum  infected dogs using  Leishmania -derived recombinant antigens.    METHODS    Leishmania  derived rK39-, rK28-, rKR95-based enzyme-linked immunosorbent assay (ELISA) was standardized using symptomatic and asymptomatic  L. infantum -infected dogs. Then 2,530 samples from inquiry in endemic areas for VL were evaluated and the results compared with recommended assays by the Brazilian Ministry of Health (MH algorithm). Further samples from a cohort of 30 dogs were searched.    FINDINGS   For rK39-, rK28- and rKR95-ELISA the sensitivity was around 97% and specificity 100%. The positivity of these three ELISA in the inquiry samples was 27-28%, around 10% higher than the assays currently in use. When cohort samples were searched, we observed likely false-negative results (> 65%) with supposedly negative samples that turned positive six months later with the assays in use (MH algorithm).    MAIN CONCLUSIONS   For the diagnosis of  L. infantum -infected dogs, rK39-based ELISA showed better diagnostic performance than other assays in use in Brazil and worldwide.

Validation of ELISA with recombinant antigens in serological diagnosis of canine Leishmania infantum infection