Ensuring optimal blood transfusion practices relies on a robust expertise base that is indispensable across various professional fields. Recognizing this imperative, the current study aims to assess the knowledge levels of healthcare personnel and enhance transfusion quality through targeted continuing training initiatives. The preliminary survey was based on an anonymous questionnaire and we used correct response rate (CRR) as the main parameter evaluating the baseline level of knowledge. Then blood transfusion education sessions which focused on transfusion shortcomings were carried out. Finally, we assessed the impact of this training on the baseline education. From the total of 789 questionnaires distributed, we garnered 331 responses 23 from healthcare staff. The overall CRR for questions related to transfusion procedure knowledge was 33%. Notably, the per-transfusion step exhibited the highest control with CRR of 55% while noticeable gaps were observed in the pre-transfusion (CRR = 22%) and post-transfusion (CRR = 13%) phases. Profession wise variations in CRR were evident, with nurses recording the lowest percentage (CRR = 29%) compared to physicians (CRR = 39%) and technicians (CRR = 34%). Substantial differences were observed in the interpretation of Ultimate Control in the Patient's Bed (UCPB) among professions, especially affecting ABO identity, compatibility, and incompatibility cases. Following the blood transfusion training, attended by 105 participants, only 99 participants responded to the questionnaire post-training, expressing high satisfaction with the covered modules (80%). The knowledge enhancement encompassed all transfusion phases and the interpretation of clinical cases, notably ABO compatibility. Overall, there was a significant improvement in CRR from 33 to 53% (p < 0.001). This study demonstrates that the evaluation of knowledge should be carried out on a continuous manner in order to detect gaps in the professional sector and to ensure the effectiveness of training through well-targeted educational programs. The online version contains supplementary material available at 10.1007/s12288-024-01778-y.