The synanthropic behavior of marsupials of the genus Didelphis in endemic areas of leishmaniasis suggests that these animals may play an important role in the epidemiology of this infection. The aim of the present study was to detect Leishmania spp. DNA in Didelphis albiventris (white-eared opossum) and Didelphis aurita (big-eared opossum) living in forested and peridomestic areas of northeastern Brazil. Blood samples were collected from 25 animals (23 D. albiventris and 2 D. aurita) by cardiac puncture and then analyzed via PCR. Leishmania (Viannia) braziliensis DNA complex was detected in 4 (16%) of the 25 samples, whereas no samples scored positive for Leishmania (Leishmania) donovani complex. Our data suggest that D. albiventris participates in the wild enzootic cycle of the L. (V.) braziliensis complex. Therefore, humans living in this area are at risk of infection, because Didelphis species are in constant movement between wild and urban environments.