Background and Objectives: The prevalence of Helicobacter pylori infection is decreasing in the Western world, while remaining high in developing countries. There is limited up-to-date information about the prevalence of H. pylori in Central and Eastern Europe. The aim of our study was to assess the seroprevalence of H. pylori and its trend over the past 25 years among students of the Lithuanian University of Health Sciences (LUHS) and to assess its relation to dyspeptic symptoms. Materials and Methods: In the years 1995, 2012, 2016 and 2020, students from Medical and Nursing Faculties of LUHS were tested for the presence of antibodies against H. pylori by performing serological tests from finger capillary blood. In addition, in the years 2012, 2016 and 2020, the students completed a gastrointestinal symptom rating scale (GSRS) questionnaire in order to assess dyspeptic symptoms. The study population consisted of 120 students in the year 1995 (mean age—21.3 ± 1.0 years), 187 students in the year 2012 (mean age—22.4 ± 0.7 years), 262 students in the year 2016 (mean age—20.4 ± 1.0 years) and 148 students in the year 2020 (mean age—20.4 ± 1.7 years). Results: The seroprevalence for H. pylori was positive in 62 (51.7%) students in 1995, in 57 (30.4%) students in 2012, in 69 (26.3%) students in 2016 and in 21 (14.2%) students in 2020. The statistically significant difference was found between all study years, except between 2012 and 2016. There were no significant differences in frequency and intensity of upper dyspeptic symptoms between H. pylori positive and negative students. Conclusions: Over the last 25 years the seroprevalence of H. pylori among students of LUHS has decreased significantly. No consistent differences in dyspeptic symptoms among H. pylori positive and negative subgroups were found.

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