Background and aim: Occult hepatitis B virus (HBV) infection (OBI) is viewed as a serious health hazard since it can lead to the advancement of acute hepatitis B virus infection, the emergence of cirrhosis, and the development of hepatocellular cancer (HCC). This study's goal was to find out how prevalent OBI was among hemodialysis (HD) patients in the Minia governorate of Upper Egypt. Methods: This study included 100 patients with end-stage renal disease getting regular hemodialysis and had negative HBV surface antigen (HBsAg) testing. ELISA was used to test sera for the presence of antibodies against HBsAg (anti-HBs) and antibodies against HBV core antigen (anti-HBc). Nested PCR with polymerase (pol) gene-specific primers and real-time PCR with surface (s) gene-specific primers were both used to detect HBV DNA. Results: Anti-HBs and anti-HBc antibodies were found in 41 and 48 % of total samples, respectively. In addition, 52.1% of anti-HBc positive patients, were positive for anti-HCV antibodies. Out of 48 anti-HBc positive samples, 33 (68.75%) samples were positive for HBV DNA. HBV DNA was shown to be significantly associated with anti-HCV antibody-positive samples (P = 0.043). Amongst samples that tested positive for anti-HBc with or without anti-HCV antibodies, there was no significant difference in ALT levels (P=0.604). Conclusion. The application of anti-HBc testing to identify OBI is a superior method to raise security in hemodialysis facilities. We recommend as well using molecular methods such as nested PCR and real-time PCR to detect HBV DNA among HD patients.