eng Oriental Scientific Publishing Company Biomedical and Pharmacology Journal 0974-6242 2025-12-30 18 4 3154 3163 10.13005/bpj/3324 69343 article Safia Khalil Ali 1 Elshazali Widaa Ali 2 Department of Hematology, Faculty of Medical Laboratory Sciences, Omdurman Islamic University, Khartoum, Sudan Department of Medical Laboratory Sciences, College of Applied Medical Sciences, University of Bisha, Bisha, Saudi Arabia Department of Hematology, Faculty of Medical Laboratory Sciences, Al Neelain University, Khartoum, Sudan β-thalassemia major (βTM) is characterized by severe anemia that necessitates regular transfusions, leading to iron overload. Mutations in the HFE gene have been reported to affect iron homeostasis and may exacerbate iron accumulation in patients with thalassemia. This study aimed to screen for HFE gene C282Y and E277K mutations in Sudanese children with βTM to assess their impact on the severity of iron overload. A total of 76 children with βTM were enrolled in the study. Genomic deoxyribonucleic acid was extracted from peripheral leukocytes and analyzed for the C282Y and E277K mutations in the HFE gene using polymerase chain reaction amplification and Sanger sequencing. Serum ferritin (SF) levels were determined using a biochemical analyzer to assess the severity of iron overload. The C282Y mutation was absent in all participants, whereas the E277K mutation was identified in 2 (2.6%) patients in a heterozygous state.  Furthermore, an intron 3 variant (rs807209) was detected in 16 (22.3%) patients within the same amplicon. Although the mean SF level was lower in patients with the E277K mutation and higher in those with the intron 3 variant than in those carrying the wildtype allele, the differences were not statistically significant (P-values = 0.140 and 0.164, respectively). In conclusion, the HFE C282Y mutation was not detected among Sudanese children with βTM. The E277K mutation was rare (2.6%) and was not significantly associated with the severity of iron overload. Similarly, the intron 3 variant (rs807209) was present in a proportion of patients (22.3%) but had no significant impact on the severity of iron overload. https://biomedpharmajournal.org/vol18no4/hfe-gene-c282y-and-e277k-mutations-as-possible-genetic-modulators-of-iron-overload-severity-in-transfusion-dependent-%ce%b2-thalassemia-major/  β-thalassemia major C282Y Mutation E277K mutation  HFE Gene Iron Overload