Objective of the study: The objective of this study was to determine the seroprevalence of Epstein-Barr virus (EBV) among children and adults based on age, sex, and serological profile. Study design: The study was designed as a retrospective investigation. Participants and methods: The data were collected from patients on an outpatient basis or during hospitalization. The samples were sent for microbiological testing. The evidence of the EBV infection was obtained using standard serological methods, including chemiluminescent immunoassay and electrochemiluminescent immunoassay. Results: The results showed that EBV seroprevalence increases with age. After the age of 20, there is a significantly higher number of patients with past EBV infection, with the majority of those with past infection being over 50 years old. A statistically significant difference was observed between men and women in the acute phase of infection, with a higher number of women having past infections. Conclusion: Seroprevalence increases proportionally with age, and as age progresses, there are fewer negative cases, but also more individuals with past infections or reactivations. This result was expected, given the previous research on EBV and the fact that 95 % of the world’s population has been in contact with the virus at least once. Children under the age of 10 are mostly negative or have an acute infection, likely due to close contact with other children during play.