Background: Beta-thalassemia stands as a prevalent variant of thalassemia among the children, presenting a substantial healthcare challenge necessitating specialized attention for both the affected children and their families. Therefore, this study aimed to investigate the effect of a stepwise program on mothers' care of their children with beta thalassemia. A pre-experimental design was employed for this study at the hematology outpatient clinic affiliated to Assiut University Children Hospital. The study involved a purposive sample comprising 115 mothers of children diagnosed with beta-thalassemia. Data collection utilized a structured interview questionnaire comprising four tools: the first part the children's characteristics and medical history, while the second part focused on the mothers' personal information. The subsequent tools assessed the mothers' knowledge, practices, and attitudes towards the management of beta-thalassemia. Findings: Post the implementation of the program, there was a notable enhancement observed in the mothers' knowledge, practices, and attitudes, with scores shifting from 12.0522±1.76140 to 23.1304 ± 6.05461, 9.0696 ± 3.66281 to 13.4087 ± 4.48955, and 2.9826 ± 1.41411 to 4.0957 ± 1.20652, respectively. Conclusion: The findings underscore the considerable efficacy of a stepwise program in augmenting mothers' knowledge, practices, and attitudes concerning the beta-thalassemia. It is recommended that the adoption of a stepwise program for mothers of children with thalassemia be implemented across all healthcare facilities catering to such patient populations.