Background: Health care associated infections and emerging multi drug resistance in nosocomial pathogens is perceived as a serious public health threat with grievous concerns. Hand hygiene if practiced properly is cheapest, simplest and most effective tool in tackling this problem. The objective of this study was conducted to assess levels of knowledge, attitude and practice in various aspects of hand hygiene in undergraduate nursing students in the study area for identifying gaps for planning necessary corrective measures. Methods: A cross sectional study involving self-administered pre-structured anonymous questionnaires administered to all undergraduate nursing students (182) enrolled at Shalmar Nursing College Lahore, Pakistan. Results: Most of the study participants exhibited moderate levels of knowledge and practice with marginal difference while attitude were found to exhibit a remarkably higher, difference being statistically significant. Conclusions: This study stresses upon the growing need for prompt interventions at institutional level for addressing the gaps evident from the study.