Background:Overall, many patients get affected by Hospital Associated Infection (HAIs) every year. Hand hygiene (HH) has been distinguished as the absolute most significant component in preventing HAIs. The point of this study was to decide the knowledge in regards to hand cleanliness among health care personnel, and recognizing the loopholes in their insight. Objective: This study aims to explore knowledge regarding hand hygiene among health care personnel. Methodology: This is a cross sectional study in Surat among 174 medical and paramedical participants selected from New Civil Hospital by Purposive Sampling. Data collection was done by digital form asking to fill up the WHO hand hygiene questionnaire for health care workers during the month of August - September, 2022. Univariate analysis was done followed by Chi square test to establish associations. Results: Mean age of population was 24.9±5.6 years. Knowledge score was classified into good (≥75%), moderate (50-75%) and poor (<50%). Majority of them had moderate knowledge (62.2%). The routine use of alcohol was significantly associated with receiving formal training in last 3 years (̒χ2:15.890, DF=1, p=0.000). Conclusion: Despite training, paramedical staff have just moderate comprehension of hand hygiene, although medical students, on the contrary, have great grasp.