Background: Iron overload is a principal reason for morbidity and death in transfusion-dependent beta-thalassemia cases. Erythroferrone hormone (ERFE), a member of tumor necrosis family- alpha (TNF-α) superfamily produced by erythroblasts and stimulated by endogenous or exogenous erythropoietin (EPO), Under these conditions, ERFE inhibits the formation of hepcidin, restoring the functionality of ferro-portin, that is accountable for enhancing intestinal iron absorption and mobilizing iron reserves. Objective: The aim of the currents study is to analyze the role of ERFE in children suffering from beta thalassemia major. Patients and methods: A prospective case-control study was conducted at Tanta University Hospital during the period from March 2021 to November 2021. The study included 40 children previously diagnosed with beta thalassemia major, and 40 healthy children matched in sex and age as a control group. Serum ERFE was calculated utilizing enzyme linked immunosorbent assay (ELISA) technique. Results: Serum ERFE level was significant increase in the beta thalassemia patients' group than control group. In the patients group, serum ERFE was higher in non spleenectomized patients, and in patients receiving blood transfusion more than once/ month, also, ERFE was higher in those with serum ferritin >1000 ng/ml. There is significant positive correlation between ERFE level and serum levels of ferritin and transferrin saturation (T.SAT %). Receiver Operating Characteristic (ROC) curve analysis demonstrated that ERFE level above 1.6 ng/ml is the cutoff value indicating iron overload with high diagnostic efficacy. Conclusion: ERFE is apossible diagnostic tool in predicting iron overload with sensitivity 95%, specificity 62 %, and accuracy 78%.