Background/aim: The aim of the present study was to determine the seroprevalence of Borrelia burgdorferi sensu lato in the city center and the province of Bolu, Turkey. Materials and methods: A stratified sampling method was used to determine the study population. A total of 196 blood samples were collected. A questionnaire was completed by each participant in the study. ELISA was performed and positive serologic results were confirmed using western blotting. Data were analyzed statistically using the chi-square test. Results: Seropositivity rates of B. burgdorferi IgM and IgG were determined as 14.8% (29/196) and 13.7% (27/196) respectively by ELISA. A total of nine serum samples (4.6%) were found positive for IgG, and seven samples (3.8%) were positive for IgM according to western blotting. Seropositivity rates were found to be higher in people living in rural areas (11.1%), in women (8.3%), in people who were illiterate (10.0%), in people engaged in agriculture (10.8%), and in the age group of 40-60 years (10.0%). No statistically significant difference was found between seropositivity rates and survey data. Conclusion: The seroprevalence of Lyme disease was determined in our region. Detection of endemic regions of Lyme disease with determination of seropositivity rates will increase the awareness among clinicians about this disease.