Background &Objectives: The Epstein-Barr Virus (EBV( is related with various diseases including infectious mononucleosis, Burkitt's lymphoma, Hodgkin's lymphoma, nasopharyngeal carcinoma and post-transplant lymphoprolifrative disorders. The aim of this study was to characterize the association between EBV and Hodgkin's lymphoma through EBERs in situ hybridization (EBER-ISH) in Iranian patients. Materials &Methods: In this study, 43 Hodgkin's lymphoma tissue samples were selected from formalin-fixed paraffin embedded blocks and analyzed by EBERs in situ hybridization. Data analyzed by SPSS16 statistical software, Fisher's exact test and Mantel-Haensel significant level 0.05. Results: 43 Hodgkin's lymphoma patients were 29 (67%) male samples and 14 (33%) female samples. In 20 (47%) samples Epstein-Barr virus was present. The positive cases included 13 samples male and 7 samples female. Fisher's exact test showed statistically no significant difference between sex and subtypes. Age distribution of relation of Hodgkin's lymphoma and EBV virus were 75% (12 of 16) in the age group of 1-14 years, 22% (5 of 23) in the age group 15-49 years and 75% (3 of 4) in the age group over 49 years, respectively. Fisher's exact test showed statistically significant difference between 1-14 and 15-49 age group years (p-value: 0.003). Conclusion: Results shown higher presence rate of Epstein-Barr virus in Hodgkin's lymphoma specimens of children and older adult. This pattern is similar to other developing countries.