The aim of this study was to detect possible differences between farmed and wild-living raccoon dogs. Analysis of polymorphism in 15 microsatellite sequences led to the conclusion that raccoon dogs raised on Polish farms and wild raccoon dogs living in Poland are two genetically distinct groups of animals. Wild Polish raccoon dogs are genetically more similar to the population of wild animals from the Kaliningrad Region than to farmed animals. The analysis of microsatellite loci showed clear genetic differences between farmed and wild-living populations of raccoon dog, despite only 50 years of isolation of the two groups of animals. The farmed population was characterized by higher genetic variation than the wild-living population. On the basis of the analyses three microsatellite loci (INU014, Ren13J22 and Ren41D20) were proposed for determination of the origin of animals that have escaped from farms.