Background: Hand washing is the most effective way to stop bacterial transmission and lower the risk of infections related to medical care. Hospital acquired infections (HAI) are known to transmit from patient to patient and within the healthcare environment most frequently from healthcare personnel. It has been proven that healthcare workers' hand hygiene practices are an effective way to lower the incidence of hospital acquired infections. Aim: A study to assess Knowledge practice and attitude regarding hand hygiene among health care professionals. Methods: The study used a cross-sectional research design. The New Najran General Hospital served as the study's location and to choose the 100 samples, a consistent sampling technique was applied. The WHO's hand hygiene questionnaire for healthcare workers was utilized as the research instrument for the study to evaluate healthcare professionals' knowledge, behavior, and attitudes toward hand hygiene. Statistics, both descriptive and inferential, were used to calculate the results. Results: The knowledge, practice, and attitude on towards hand cleanliness was adequate and the practice score was 3.72 with a standard deviation of 0.514, and the overall mean total knowledge score was 8.36 with a standard deviation of 1.599. The attitude rating was 2.48, with a 1.176 standard deviation. Conclusion: The results of this study showed that healthcare workers at New Najran General Hospital have acceptable knowledge, attitudes, and hand hygiene practices. We advocate for the supply of these necessities as well as teaching sessions to help caregivers and patients understand the value of hand washing