The current study aims to determine the relationship of ß-thalassemia and its types (major, minor, intermedia) with ABO and Rh blood groups in Lahore, Pakistan. It is an observational study in which ABO and Rh blood grouping of 711 β-thalassemia patients was performed (with their consent). Blood grouping was performed through forward and reverse grouping using tube method. The observed ABO blood group pattern in both genders was O>B>A>AB. It was found that the overall frequency of O and A blood groups was higher in female patients, while the frequency of B and AB blood groups was higher in male patients, although the difference was non-significant. However, a significant difference was observed in the relationship between gender and Rh factor. Most patients of β-thalassemia suffered from the type β-thalassemia major (48%), followed by patients suffering from β-thalassemia minor (44%) and β-thalassemia intermedia (8%). Overall, patients with AB blood group mostly suffered from β-thalassemia major, while patients with β-thalassemia minor and β-thalassemia intermedia had the highest frequency of blood group B. It was concluded that ß-thalassemia patients have a higher frequency of O+ blood group and the lowest frequency of AB- blood group. Moreover, a significant difference was seen between male and female patients with respect to Rh blood group. However, no significant difference (p > 0.05) was found in the distribution of ABO and Rh blood groups corresponding to β-thalassemia types.