Objective To understand the implementation status of Regulation of disinfection technique in healthcare settings (WS/T367) (hereinafter referred to as regulation of disinfection), find out the main problems and weak links in the disinfection management, and provide scientific basis for further strengthening the disinfection management of healthcare settings. Methods A multi-stage stratified sampling method was adopted to determine the surveyed subjects, formulation and revision of hospital disinfection management system, management of disinfection products, supervision of disinfection work, disinfection-related training, cleaning and disinfection process were investigated. Results A total of 256 healthcare settings were investigated, after the issuing of regulation of disinfection, 94.14% (n=241) of healthcare settings formulated or revised disinfection management system according to the actual situation; 93.75% (n=240) of healthcare settings reviewed, backuped and stored relevant certificates of disinfection products; 91.41% (n=234) of healthcare settings regularly supervised the disinfection work, as well as summarized, analyzed and fed back in time; 93.75% (n=240) of healthcare settings carried out disinfection-related training, and 88.67% (n=227) of healthcare settings carried out the correct routine reuse process of medical instruments. Conclusion The issuing of regulation of disinfection standardized the disinfection management of healthcare settings. Disinfection process of reusable instruments in healthcare settings has been basically standardized, correct rate of cleaning and disinfection process in primary healthcare settings is obviously lower than that in hospitals, which is the focus of disinfection management in the future.