Aphanomyces astaci is a fungus-like oomycete agent responsible for an illness called crayfish plague, reaching 100% mortality in infected animals. Therefore, the aim of the work was to detect and estimate the rate of infection of spiny-cheek crayfish (Orconectes limosus) by A. astaci in selected water reservoirs of north-western Poland, as this crayfish is described as a main cause of crayfish plague. The material for the study were 54 spinycheek crayfish individuals from 3 sites in Poland: Trzebiocha River, Lake Sominko and Lake Dąbie. A total of 162 samples (muscle samples were taken from abdomen, legs and carapace) were taken and used for DNA extraction followed by PCR and bidirectional sequencing of 5.8S ribosomal RNA gene. The electrophoretic separation of the PCR products confirmed the presence of A. astaci in 17 samples (Trzebiocha River and Lake Dąbie). Lake Sominko proved to be a zone free of the investigated pathogen. The collected information on the presence or absence of A. astaci in the investigated reservoirs might be used for restocking purposes.