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Background: The hepatitis B virus (HBV) is one of the major causes of chronic liver disease. From the perspective of hospital workers (HWs), employees are at risk of hepatitis B infection because of occupational exposure. Apart from this occupational risk, health professionals may still be affected by HBV, depending on the epidemiological characteristics of the country and geographical region they live in. Objectives: This study aimed to determine HBV, HCV, and HIV seroprevalence among HWs using data obtained from 21 hospitals located in six geographical regions in Turkey and the Turkish Republic of Northern Cyprus. Methods: The study was designed as a retrospective, multicentre, descriptive study. Twenty hospitals from Turkey and one hospital from the Turkish Republic of Northern Cyprus were involved in the study. The variables of the study were vaccination status against HBV and hepatitis A and HBsAg, anti-HBs, anti-HBcIgG, anti-HAVIgG, anti-HCV, anti-HDV, and anti-HIV serology results belonging to the previous year. Results: Women constituted 58.9% (n = 5,622) of the HWs included in the study. The mean age was 36.3 +/- 9.09 years (min =18, max = 72). In terms of occupation, 42.5% (n = 4, 064) were nurses/health officers, and 24.8% (n = 2365) were physicians. HBsAg seroprevalence was found to be 1.8% (n =169; 95% CI = 1.5% - 2.0%), while anti-HBs seropositivity was 75.7% (n = 7,234). About 7.3% (n = 701) had natural immunity to hepatitis B. About 21.6% (n = 2,066) of the HWs did not receive hepatitis B vaccine. Conclusions: This study is the first study involving a large sample size from different locations of Turkey. According to the results, hepatitis B and hepatitis A vaccines should be administered to susceptible individuals and HWs. WOS:000534475500002 2-s2.0-85085751514