Introduction: Blood transfusion is a critical intervention in neonatal care, particularly for premature infants, to address complications such as anemia of prematurity and hemorrhagic shock. This study aimed to evaluate blood transfusion practices and outcomes in neonates at Kamenge Teaching Hospital, Burundi, addressing a gap in the literature specific to this region. Materials and Methods: A prospective, observational study was conducted from January to August 2024, including 48 newborns who met the eligibility criteria and underwent blood transfusion. Data were collected using pre-established forms and analyzed using Microsoft Word 2010 for text entry, and Excel 2010 and SPSS version 25 for data analysis and processing. The study was approved by the Ethics Committee of Kamenge Teaching Hospital, and informed consent was obtained from the parents or legal guardians of each participant. Results: The study found a higher proportion of female newborns (54.2%) and those from rural areas (64.6%). Prematurity was the leading cause for hospitalization (42.7%), and the majority had moderate anemia (62.5%). The majority of transfusions were well-tolerated, with a low incidence of transfusion-related accidents (6.3%). Favorable clinical evolution was observed in 83.3% of cases, while the mortality rate was 16.7%. Conclusion: The study underscores the critical need for optimizing blood transfusion practices and neonatal care in Burundi. Recommendations include reviewing transfusion protocols, strengthening management measures for preterm infants, expanding preventive health measures in rural areas, and investing in the training and education of healthcare providers. The study also highlights the importance of ongoing monitoring and further research to improve neonatal health outcomes in low-resource settings.