Introduction: Hand Hygiene is an essential method to preclude infections in all healthcare environments. Education is essential to advance hand hygiene performance amongst nurses and nursing students. The aim is to find out the usefulness of a self-paced online hand hygiene course on the knowledge, attitude and practice of nursing students by evaluating student performance in post-tests. Methods: The study employed a quasi-experimental design in which data were collected using two questionnaires from undergraduate nursing students and they were exposed to interactive lectures and online activities related to risks, benefits, and key recommendations for hand hygiene. The following two surveys were used, Hand Hygiene Knowledge Questionnaire for Health-Care Workers; and Handwashing Assessment Inventory. Data were collected through pre-post tests. Results: Nurses reported a significant change for the better in hand hygiene comprehension, attitude, and performance of hand hygiene behaviours. The present study revealed a significant increase in hand hygiene knowledge belief, attitude, and outcomes among university students after participating in the training intervention. The majority of participants in the pretest rated their knowledge level as poor. Conclusion: Interactive training and online learning courses on hand hygiene could be used to influence the beliefs, attitudes and behaviours of students to bring about the desired change in hand hygiene practice.