Objective To understand the cognition and influencing factors of hand hygiene (HH) among healthcare-associated infection (HAI) management staff in Gansu Province, and provide reference for enhancing the special competency of HAI management staff. Methods According to geographical area, natural and human characteristics, HAI management staff in 69 hospitals in different areas of Gansu Province were selected to conduct a questionnaire and field survey on their basic condition and knowledge of HH, current situation, and influencing factors for HH knowledge were analyzed. Results Among 69 surveyed hospitals, 82.61% were secondary hospitals, 17.39% were tertiary hospitals; among 125 surveyed HAI management staff, 82.40% were full-time staff and 17.60% were part-time staff. 33.60% and 66.40% of HAI management staff had complete and incomplete knowledge of HH respectively; the knowledge about importance of HH, use of hand disinfectant, installation of HH facilities, hand washing methods, hand drying methods, HH indications, and standard for judging qualified HH in tertiary hospitals were all higher than those in secondary hospitals (all P<0.05). Multivariate logistic regression analysis showed that independent protective factors for HH cognition were working for ≥ 5 years and <10 years in HH management (OR=3.067), bachelor's degree (OR=3.331), and full-time job (OR=10.309). Conclusion HAI management staff's cognition on HH among is low, especially those who have been engaged in HH management for less than 5 years, whose educational background is secondary school or lower, and part-time personnel. It is necessary to combine with actual condition to strengthen professional competence training for HH management staff.