Background: Thalassaemias are inherited disorders of hemoglobin synthesis, with β-thalassaemia being the most prevalent and clinically significant. In Bangladesh, the burden is high, with over 7000 affected births annually and notable carrier rates of both β-thalassaemia and Hb E, especially among tribal populations. The disease arises from β-globin gene mutations, leading to imbalanced globin chain production and severe anemia. Regular transfusions are essential but pose risks, including iron overload and transfusion-transmitted infections like hepatitis C virus (HCV). HCV affects over 60% of transfusion-dependent patients, significantly increasing morbidity and mortality, yet long-term survival data and treatment outcomes remain inadequately explored. Aim of the study: This study aims to compare the hematological and biochemical parameters of HCV-infected and non-infected multi-transfused thalassaemic patients. Methods: This cross-sectional observational study was conducted over seven months (February–August 2015) at the Department of Paediatric Haematology and Oncology, Dhaka Shishu Hospital, Bangladesh. Sixty β-thalassemia major patients aged 4–18 years receiving regular transfusions were enrolled, including 11 anti-HCV seropositive and 49 seronegative cases. Participants were recruited from the thalassemia clinic and transfusion unit. Data were collected through interviews, clinical exams, and structured checklists. Laboratory investigations included ELISA for anti-HCV, hematological and biochemical analyses, and hepatic ultrasonography. Statistical analysis was performed using SPSS v16.0 with Chi-square and Student’s t-tests; p < 0.05 was considered statistically significant. Result: HCV-positive patients had a significantly higher mean age (11.6±3.8 vs. 9.3±4.6 years; p=0.032) and body weight (24.6±8.8 vs. 15.8±5.5 kg; p=0.001) than HCV-negative patients. Jaundice (p=0.043), abdominal pain (p=0.008), hepatomegaly (p=0.011), and splenomegaly (p=0.035) were significantly more prevalent in the HCV-positive group. Splenomegaly and hepatomegaly sizes were also larger. No significant differences were found in sex, socioeconomic status, hemoglobin, HBsAg status, or most hematological and biochemical markers. However, serum bilirubin was significantly elevated in HCV-positive patients (2.2±0.5 vs. 1.5±1.1 mg/dL; p=0.044), indicating greater hepatic involvement. These findings suggest a higher disease burden among HCV-infected individuals. Conclusion: HCV infection in multi-transfused thalassaemic patients is linked to greater liver involvement, including jaundice, hepatomegaly, and elevated bilirubin. Older age suggests cumulative transfusion risk. Despite similar hemoglobin levels, liver disease burden is higher in HCV-positive individuals, highlighting the need for early screening, safer transfusions, and targeted liver monitoring.