Two short-tailed opossum species, Monodelphis glirina and M. touan, occur in sympatry in an area of eastern Amazonia. Habitat structure and resource availability may influence habitat use and, consequently, species distribution, detectability, occupancy, and abundance. We evaluated occupancy and detectability of the species M. glirina and M. touan in the Carajás National Forest to answer the following questions: (1) Do both species occur in canga and forest habitats? (2) Do detectability and occupancy of M. touan and M. glirina differ in areas of canga and forest? (3) Does the presence of one species affect the detection of the other? We undertook surveys at 50 sampling sites (26 in canga and 24 in forest habitat). In addition, we developed co-occurrence models to test the relationships between occupancy and detection of M. touan in the presence or absence of M. glirina. We captured 693 individuals of M. glirina (587 in canga and 106 in forest) and 112 of M. touan (only one individual captured in canga). Occupancy by M. glirina was positively influenced by superficial rock cover and litter depth, while detectability was negatively influenced by canopy cover. Occupancy by M. touan was influenced positively by canopy cover, number of fallen trunks on the ground, and litter depth (Ψ = 0.315). Data from forest sites where M. touan and M. glirina occurred more often corroborated our third hypothesis, that detectability of M. touan is low when M. glirina is present. Our results highlight the existence of habitat preference by the two species of Monodelphis.