Infections Acquired in healthcare setups are among the major causes of death and increased morbidity among hospitalized patients especially in developing countries. Healthcare-Associated Infections (HAIs) significantly cause patient mortality and morbidity. Hand hygiene is one of the most effective, cheapest ways to prevent Nosocomial infection. Health care worker’s hands are the most usual type of vehicle for transmission of healthcare-associated infections from patient to patient and within health care environment.

Aim: To determine the knowledge and practice levels among Al-Imam Al Hussein medical city nursing staff. Materials and methods: Cross-sectional study was conducted at Al-Imam Al Hussein medical city, Karbala from November 2022- March 2023, a total of 128 subjects were participated. The target population were nursing staff and collection of data was through structured and analyzed using a statistical package for social sciences.

Results: About 40.6% of nursing staff had good level of hand washing knowledge, 57% fair level and 2.3% poor level. Also, about 71.9% of the nursing staff had good level of hand washing practice and 28% had a fair level.

Conclusions: The hand washing knowledge and practice are improving among nursing staff at Al-Imam Al Hussien Medical City. We recommended continuous hand hygiene training programs, workshops and saving hand hygiene supplies.