Background and Aims: Blood and components are more frequently used in surgery and non-surgical procedures. In medical procedures blood transfusion is important and needs adequate expertise and practice, thus adequate knowledge in healthcare workers of this procedure is essential. Materials and Methods: This descriptive study is designed to assess the knowledge of healthcare workers about proper methods of blood transfusion, and how to promote their knowledge for proper performance if their knowledge is inadequate. Data were collected with aimed questionnaire and analyzed by statistics software. Result: The study population mainly comprised 122 healthcare workers (HCWs). The main findings from this study showed that 26.2% of healthcare workers (HCWs) had low-level knowledge, 22.1% moderate and 51.6% acceptable knowledge. We did not find any significant correlation between knowledge of HCWs and years in profession, participation in training, number of blood transfusions per day, age, gender, etc. (P < 0.05). Discussion and Conclusion: Results strongly emphasized the need for a curriculum to promote knowledge of HCWs about blood transfusion because we found low and moderate level of knowledge in approximately half our samples and on the importance of blood transfusion procedure, suggesting that more attempts should be made to build up knowledge about blood transfusion.