We report a new locality for Monodelphis handleyi, a rare short-tailed opossum species, previously known only from its type locality, in Loreto, northeastern Peru. One adult male was collected using pitfall trap disposed in Humaitá Amazonian savanna of southern Amazonas state, Brazil. Voucher specimen had their identification confirmed by molecular data (mitochondrial gene Cytochrome b) and morphological comparison. We provide external and cranial measurements of this specimen and comment on its morphology. The specimen reported here represents the first record of M. handleyi in Brazil and is the second known locality for the species, and there is high level of divergence found between the two distant localities (5.1 %). This record extends the species range at least 1,200 km eastern beyond the type locality and denotes the importance of use of complementary methods for sampling small nonvolant mammals.