Background: Worldwide, medical staff adherence to hand hygiene (HH), the most cost-effective measure to decrease healthcare-associated infections (HAIs), is ~40%–60%. The infection control program (ICP) at An-Najah National University Hospital (NNUH), a tertiary-care referral teaching hospital located in Nablus, in northern Palestine, monitors HH compliance by direct observations using the WHO observation checklist. In this descriptive study, we investigated the prevalence of HH across the institution during 2017–2019. Methods: The WHO multimodal strategy to enhance HH in healthcare settings was implemented at NNUH, a tertiary-care referral hospital, in 2017. HH compliance has been measured during routine patient care by direct observation by ICP team and anonymously by other trained observers. Results are reported on monthly basis to the administration and medical team (nurses and doctors), and corrective plans to increase the compliance are discussed. Training is reinforced with ultraviolet light and fluorescent alcohol-based hand rub. Yearly, staff are engaged in HH Day activities (Figs. 1 and 2). Leadership support is constant by securing the annual budget for the HH program and the enforcement of HH policy across the setting. Results: NNUH, using the WHO Hand Hygiene Self-Assessment Framework, is currently in the advanced level (395 of 500) compared to 2017 (intermediate level, 292 of 500). Overall, HCW HH compliance increased from 44% (range, 31%–57%) in 2017 to 53% (range, 30%–72%) in 2018 and to 61% (range, 55%–66%) through October 2019. During the 3-year study period, Nursing compliance increased from 36% to 59% and to 64%, respectively, whereas the compliance for doctors increased from 42% to 56% and 58%, respectively. Regarding the missed opportunities, before patient was the most frequent cause for missed opportunity with a rate of 60% (Fig. 3). This missed opportunity was mainly related to the misuse of gloves (mainly among nurses), which requires an ad hoc intervention. Conclusions: Implementation of the WHO HH strategy is feasible and effective in low-income countries and leads to significant improvements in compliance. Periodic training, personnel engagement, and leadership are key factors of HH improvement in our setting.