Although ferrets are becoming increasingly popular as companion animals, their population in households is still far lower compared to cats or dogs. This results in a much smaller number of ferret specialists, and thus poorer diagnosis of various diseases, including the Aleutian disease. Aleutian disease is a slowly progressing viral disease which can cause different symptoms in these animals. The virus can also cause symptoms in different species of animals, but in the case of ferrets, there is relatively less information on about both the prevalence and symptoms of this disease. Therefore, the aim of this study was to determine the presence of antibodies and the virus itself in ferrets from two regions of Poland. Blood samples and rectal swabs were obtained from 61 domestic ferrets from Mazowieckie and Dolnośląskie voivodships. The presence of antibodies was determined using serological methods and real-time PCR analysis was performed to determine presence of viral DNA. Serological analyses demonstrated that 49% (n = 30) of the ferrets had antibodies against Aleutian disease virus (ADV). No relationship was observed between the prevalence of antibodies and age, sex, habitual residence or origin of ferrets. The real-time PCR did not confirm DNA of the ADV in any of the blood and rectal swab samples. Obtained results suggest that ADV circulates in the analyzed population of ferrets, therefore further studies in this direction should be carried out.