Background:Epstein-Barr virus (EBV), a gamma-1 herpes virus, is a lifelong asymptomatic infection that is carried by the vast majority of human populations. However, it appears etiologically related to several pre-malignant lymphoproliferative diseases. It often accompanies chronic liver disease and cirrhosis. EBV acts as a catalyst for virus C, demonstrating a connection between Hepatocellular carcinoma (HCC) and EBV virus. Objective: The aim of the current study is to evaluate the EBV prevalence in HCC patients, and whether it is contributed to the development of HCC by working synergistically with hepatitis C virus.Patients and methods:The study was conducted and included 90 HCC patients, 45 chronic liver disease patients, and 45 controls. Full history was taken, and clinical data of the patients were noted. Blood samples were drawn, and the following lab tests were performed: CBC, prothrombin time, alpha-fetoprotein, and liver function tests (AST, ALT, albumin, bilirubin). EBV IgM and IgG were done by ELISA technique. DNA extraction and EBV detection by PCR were performed for those who had positive EBV IgM. Results: EBV IgM antibodies was higher among HCC group compared to CLD group, while there was no difference in EBV IgG antibodies between the two groups. According to EBV detection by PCR, no difference was observed between the HCC and CLD groups in samples that tested positive for EBV IgM. Conclusion:EBV may have a role in development of HCC.