Objective To explore the pathogen contamination and cost-effectiveness of hospital bed-side curtains under different cleaning frequencies. Methods Fourteen departments in a hospital with relatively higher incidence but smaller fluctuation of incidence of infection from January to June 2023 were selected. Nine wards in each department were selected, and two curtains were selected from each ward. According to the cleaning frequency, wards were divided into low, medium and high frequency groups. The qualified rate of environmental hygienic detection on pathogens on the bed-side curtains and bacterial contamination on the surface of curtains after 6 months were comparted, cost-effectiveness analysis was conducted. Results The qualified rate of environmental hygienic detection of bed-side curtains was the highest in high frequency group, at 88.10%. Bacterial counts on the upper, middle, and lower segments of the bed-side curtains in the high frequency group were all lowest (P<0.05). Within 6 months, the incidence of healthcare-associated infection (HAI) in patients in the high and the medium frequency groups (0.16% and 0.25%, respectively) were both lower than 0.85% in the low frequency group (χ²=5.653, 3.915, respectively, both P<0.05). Cost-effectiveness analysis showed that for every additional unit of qualified rate for the high and medium frequency groups, the costs increase by 124.51 and 44.10 Yuan, respectively. Conclusion The increase of cleaning frequency can effectively reduce bacterial contamination of bed-side curtains, decrease the risk of HAI, and improve the qualified rate of environmental hygiene of bed-side curtains in the wards. Cleaning once every 2 months can achieve better cost-effectiveness.