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This study analyzed 1464 surface samples from 136 operating rooms at 23 hospitals.When used after manual cleaning, PX-UV reduced contamination on operating room surfaces.PX-UV can be an adjunct to manual cleaning processes.Manual cleaning and disinfection of the operating room (OR) environment may be inadequate due to human error. No-touch technologies, such as pulsed-xenon ultraviolet light (PX-UV), can be used as an adjunct to manual cleaning processes to reduce surface contamination in the OR. This article reports the cumulative results from 23 hospitals across the United States that performed microbiologic validation of PX-UV disinfection after manual cleaning.We obtained samples from 732 high-touch surfaces in 136 ORs at 23 hospitals, after manual terminal cleaning, and again after PX-UV disinfection (n = 1464 surface samples). Samples were enumerated after incubation, and the results are reported as total colony-forming units (CFU).The average CFU after manual cleaning ranged from 5.8 to 34.37, and after PX-UV, from 0.69 to 6.43. With manual cleaning alone, 67% of surfaces were still positive for CFUs; after PX-UV disinfection, that number decreased to 38% of all sampled surfaces—a 44% reduction. When comparing manual cleaning to PX-UV, the reduction in CFU count was statistically significant.When used after the manual cleaning process, the PX-UV device significantly reduced contamination on high-touch surfaces in the OR.