Daily cleaning and disinfecting of non-critical surfaces in the patient-care areas are known to reduce the occurrence of health care-associated infections. However, the conventional means for decontamination of housekeeping surfaces of sites of frequent hand contact such as manual disinfection using ethanol wipes are laborious and time-consuming in daily practice. This study evaluated a newly developed portable pulsed ultraviolet (UV) radiation device for its bactericidal activity in comparison with continuous UV-C, and investigated its effect on the labor burden when implemented in a hospital ward. Pseudomonas aeruginosa, Multidrug-resistant P. aeruginosa, Escherichia coli, Acinetobacter baumannii, Amikacin and Ciprofloxacin-resistant A. baumannii, Staphylococcus aureus, Methicillin-resistant S. aureus and Bacillus cereus were irradiated with pulsed UV or continuous UV-C. Pulsed UV and continuous UV-C required 5 and 30 s of irradiation, respectively, to attain bactericidal activity with more than 2Log growth inhibition of all the species. The use of pulsed UV in daily disinfection of housekeeping surfaces reduced the working hours by half in comparison to manual disinfection using ethanol wipes. The new portable pulsed UV radiation device was proven to have a bactericidal activity against critical nosocomial bacteria, including antimicrobial-resistant bacteria after short irradiation, and was thus found to be practical as a method for disinfecting housekeeping surfaces and decreasing the labor burden.