Objective To survey the implementation of Technique Standard for Isolation in Hospitals (WS/T 311-2009)(referred to as isolation standard) at all levels of hospitals in China, and provide scientific basis for the revision of isolation standard. Methods In 2017, 253 hospitals in 12 provinces (municipalities) in China were selected to survey the implementation of isolation measures before and after the issuing of isolation measures, including revision of isolation system, training, supervision and other management work, building layout and installation of facilities, implementation of hand hygiene, implementation of standard prevention relevant work, use of protective equipment, and implementation of occupational exposure management. Results In 2017, of 253 surveyed hospitals, 99.21% (n=251) established isolation prevention system, 97.23% (n=246) regularly guided and supervised isolation work; implementation of hand hygiene reached peak in 2009; 76.28% (n=193) of hospitals set up intestinal outpatient clinics, 79.84% (n=202) set up fever clinics, 88.54% (n=224) set up outpatient and emergency pre-examination and triage; 99.21% (n=251) carried out standard prevention relevant work; compared with 2010, the use of medical personal protective equipment increased more than double; 98.02% (n=248) of hospitals had reporting procedure and system of needle stick injury. Conclusion Issuing of isolation standard has played a promoting role in standardizing the isolation work of hospitals.