Background: Infection is the most frequent complication related to central venous catheters and usually occurs when microorganisms on the patient’s skin or health cares’ hands migrate down the catheter tract or through its hubs and subsequently colonize the catheter. Nurses are often at the frontline of central line care and the profession has the potential to significantly impact the reduction in central line associated blood stream infections by adopting preventive measures.1Aim: The aim of the study was to evaluate the effectiveness of structured teaching programmein terms of knowledge and practice regarding care bundle on prevention of central line associated blood stream infection among staff nurses. Methodology: Pre-experimental research design was adopted and Non-probability convenient sampling technique was used to collect the sample from 30 staff nurses working in intensive care units. Data obtained was analysed and interpreted in the light of the objectives using both descriptive and inferential statistics. Results: The findings of the study revealed that the mean post-test knowledge and post-test practice scores of the staff nurses working in intensive care units was significantly higher than the mean pre test knowledge and mean pre test practice scores. Also, there exists a positive correlation between post test knowledge scores and post test practice scores among staff nurses working in intensive care units. Conclusion: Thus, the research study findings indicated that the structured teaching programme was effective in improving the knowledge and practice of staff nurses regarding care bundle on prevention of central line associated blood stream infection.