Kazakhstan is being considered medium-endemic for Hepatitis B virus infection (HBV). HBV remains transmissible by direct exposure to infected blood or organic fluids. This cross-sectional study aimed to evaluate the prevalence of anti-HBcore and the risk factors impacting positive anti-HBcore markers among donors at Scientific-Production Center of Transfusiology, Ministry of Healthcare of the Republic of Kazakhstan. The samples taken from blood donors were tested for anti-HBcore, by the chemiluminescence immunoassay method on the Architect i2000SR (Abbott). In case of positive anti-HBcore, the blood samples were further tested for anti-HBs on the Architect i2000SR (Abbott). Alanine aminotransferase (ALT) indicators were tested by kinetic method on the Biosystems A25 analyzer. Statistical analysis was conducted using R software (version 4.1.1, 2021). Five thousand seven hundred and nine people aged 18-66 years included in the study, the proportion of men and women was 68.17% and 31.83%, respectively. The average age of the participants was 35.7 ± 10.57 years. The prevalence of anti-HBcore among donors was 17.2% (983). Among participants with elevated ALT (170), this marker was determined in 23%, and for donors with normal levels of ALT (5539)-17%. Participants with positive anti-HBcore scores were on average older (41.8 vs. 34.4 years, p < .001) and Kazakhs (88.7% vs. 83.0%, p < .001) by nationality than study participants with negative results of anti-HBcore. Anti-HBcore prevalence in Kazakhstan (17.2%) compared with other countries (Croatia 7%, France 7%, Germany 9%, Iran 16%, Malaysia 20%, respectively) remains above average. Given the prevalence of HBV and risk factors, it is recommended to include an additional anti-HBcore marker in the mandatory screening of donated blood in the Kazakhstan Republic and improve preventive measures to prevent HBV transmission by blood transfusions.