Few studies of population structure and genetic diversity exist for frogs in the Amazon of South America, an area renowned for exceptionally high species richness. We isolated seven highly variable tetranucleotide microsatellite loci for the neotropical leaflitter frog, Eleutherodactylus ockendeni using an enrichment method. Three of the repeats are simple, three are compound and one is imperfect. We screened all loci with 175 individuals from one geographical area in the upper Napo of Ecuador and found high polymorphism in all loci (> 14 alleles/locus). These markers are suitable for population genetics studies of E. ockendeni and perhaps other leaflitter frogs of the same genus.