Background: Hepatitis is a serious blood born infection in patients with β-thalassemia major (β-TM). There was no previous report on hepatitis prevalence in patients with β-TM in Zabol, Iran. Objectives: The current study aimed to evaluate characteristics of hepatitis in patients with β-TM visiting Imam Khomeini Hospital of Zabol, in Sistan and Baluchistan province of Iran. Patients and Methods: There were 90 patients with β-TM. General demographic and laboratory results were obtained by interviews and reviewing clinical and laboratory histories. Data for anti-HCV, HBs antigen and anti-HBs were extracted from latest archived records, and analyzed using SPSS ver. 19. Results: Males and females comprised 51% and 49% of the patients, respectively. Mean age of the patients was 14.8±7.4 years old. Anti-HCV and HBs antigen were detected in 10% and 3.3% of the patients, respectively. Significant difference was found between the mean age of anti-HCV positive (20.1 ± 3.6, 95% CI: 17.3-22.9) and negative (14.2 ± 7.5, 95% CI: 12.5-15.9) patients (P = 0.02). Mean ferritin value was measured 4702 ± 2743 ng/mL. A significant correlation was observed between serum ferritin level and age (r = 0.3, P = 0.01). Hepatic enzymes, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) had mean levels of 51.8 ± 32.8 IU/L and 58.8 ± 59 IU/L, respectively. No significant association was identified between hepatitis status and either ferritin or hepatic enzymes levels. Conclusions: Results demonstrated the relatively high prevalence of HCV infection in the patients. Applying sensitive methods to screen blood units is recommended to minimize the risk of transfusion associated hepatitis.