Objective To understand the effect of World Health Organization(WHO) multimodal hand hygiene improvement strategy on hand hygiene compliance among acupuncturists. Methods All the acupuncturists in departments (Department of Acupuncture, Department of Encephalopathy, Department of Orthopedics and Traumatology) with acupuncture programs in Xi’an Hospital of TCM were chosen in this study between September 2015 and August 2016. Based on the WHO multimodal hand hygiene improvement strategy, comprehensive measures were regulated among acupuncturists. Hand hygiene compliance and accuracy, and hand hygiene knowledge score were compared before and after the strategy intervention. Then, the effects of key strategies were evaluated. Results Overall hand hygiene compliance rate, accuracy and knowledge scores increased from 51.07%, 19.86% and 81.90±2.86 before intervention to 72.34%, 51.70%, and 98.62±2.92 after intervention (P<0.05). Hand hygiene compliance rates also increased in various occasions such as before contacting the patient, after contacting the patient, before acupuncture treatment, and before acupuncture needle manipulation (P<0.05). Conclusion Hand hygiene compliance in acupuncturists can be significantly improved by the implementation of WHO multimodal hand hygiene improvement strategy.