β-thalassemia is a common genetic disorder affecting β globin gene and itis characterized by ineffective erythropoeisis and iron overload as asecondary complication. HFE gene is considered to be a major candidatefor the gene bearing the primary defect responsible for hemochromatosisand iron overload. The aim of the study was to evaluate the influence ofH63D and C282Y mutations in HFE gene on serum ferritin and ironlevels in different genotypes of thalassemic patients and carriers. H63Dand C282Y mutations of HFE gene were assessed in 75 β-thalassemiapatients and in 25 β-thalassemia carriers by polymerase chain reaction –restriction fragment length polymorphism. In addition determination ofthe underlying genetic mutation in β- globin chain was done throughreverse hybridization technique. The highest prevalence of the underlyinggenetic mutation of β globin gene was shown in IVS 1.110 mutationfollowed by IVS 1.6. The C282Y mutation showed 100% wild typegenotype in both the patient and carrier groups while a higher wild allelicfrequency (H allele) of H63D mutation was noticed in carriers comparedto thalassemic cases (P value 0.002). A significant positive relationbetween the presence of mutant D allele of H63D mutation and thehigher ferritin levels was detected among our thalassemic patients. Thehighest levels of ferritin were encountered among homozygous mutantpatients. In conclusion, screening of H63D mutation may help to predictpatients with higher tendency of developing iron overload.