A qPCR-based strategy for differential diagnosis of visceral and cutaneous leishmaniasis in humans and dogs.

This study developed a quantitative real-time PCR (qPCR)-based strategy for the differential molecular diagnosis of visceral and cutaneous leishmaniasis in humans and dogs, using blood, skin biopsy, and conjunctival swab samples. The assay demonstrated satisfactory sensitivity and specificity, reaching 93.3% and 100% in human samples and 90.48% and 100% in canine samples, across the evaluated species (Leishmania infantum, L. braziliensis, and L. amazonensis). Detection of L. amazonensis required an adapted protocol incorporating SYTO9 intercalating dye and melting curve analysis. Although exploratory, these results indicate that the proposed method is feasible and support its potential for further validation in larger cohorts and future studies.