Background: Thalassaemia remains a major public health concern in Bangladesh, where hereditary disorders are increasingly recognized as contributors to long-term morbidity. Junior doctors play a critical role in early identification, counselling and implementation of preventive strategies. This study aimed to evaluate the level of knowledge and attitude of junior doctors toward thalassaemia and compare findings between interns and medical officers. Methods: A hospital-based cross-sectional descriptive study was conducted among 110 junior doctors (interns and medical officers) in Gonoshasthaya Samaj Vittik Medical College, Savar, Dhaka, Bangladesh, from April to June 2019. Data were collected using a pre-designed, self-administered questionnaire comprising demographic variables, 20 knowledge items and 9 attitude items. Knowledge scores were categorized as good, average, or poor. Attitude scores were classified as positive or negative. Data were analyzed using SPSS. Results: Participants were predominantly aged 22–26 years (76.4%), female (63%) and interns (72.7%). Good knowledge was observed in 34.5%, while 34.5% had average and 30.9% had poor knowledge. Positive attitudes were noted in 81.8% of respondents. Interns demonstrated significantly better understanding of consanguinity (p=0.014), prevention (p=0.011) and routine investigations (p=0.008). All participants were willing to donate blood and 80% were willing to donate bone marrow. Knowledge and attitude showed a positive, though statistically insignificant, association. Conclusion: Despite favorable attitudes toward prevention, substantial knowledge gaps persist among junior doctors. Strengthened training in genetics and thalassaemia management is necessary.