Objective To investigate healthcare-associated infection (HAI) management professionals' awareness on disinfection knowledge, explore the possible influencing factors. Methods Questionnaire with good reliability and validity was used to investigate disinfection knowledge among 135 HAI management professionals. Results HAI management professionals' scores of knowledge dimension, behavioral dimension, and total score of knowle-dge-attitude-practice (KAP) towards disinfection in tertiary hospitals were all higher than those of secondary hospitals ([11.12±2.09] vs[10.00±1.47], [10.58±1.96] vs[9.34±1.64], [25.35±3.83] vs[23.61±2.52], respectively, all P < 0.05); there was no significant difference in the score of attitude dimension of KAP in different hospitals (P=0.359). KAP score of HAI management professionals in 40-49 years group was the highest (26.33±3.04); the higher the education background, the higher the KAP score; KAP score of chief nurse or associate chief nurse (26.90±2.71) was higher than that of other professional titles; KAP score of full-time HAI management professionals (27.61±2.52) was higher than that of part-time staff; disinfection KAP score of HAI management professionals with 5-10 working years (26.47±2.63) was higher (all P < 0.05). Conclusion Awareness rate of disinfection knowledge among HAI management professionals needs to be improved, especially training on disinfection knowledge in secondary hospitals should be strengthened, it is necessary to carry out targeted intervention measures in different groups of population.