This study determined the efficacy of photocatalytic HEPA filters on microorganism disinfection in a closed-loop chamber and later applied it in an air purifier and tested its efficacy in an 8-m3 chamber and in a hospital. The photocatalytic filters were made by dip-coating a HEPA filter in a TiO2 slurry. In order to disinfect the microorganisms retained on the filter, UV-A light was irradiated onto the filter to create strong oxidative radicals which can destroy microorganisms. The findings showed that disinfection efficiency of the photocatalytic filters with high TiO2 loading was insignificantly higher than with lower loading. S. epidermidis was completely eliminated within 2 hours, while 86.8% of B. subtilis, 77.1% of A. niger, and 82.7% of P. citrinum were destroyed within 10 hours. When applying the photocatalytic filters into an air purifier in a 8-m3 chamber, it was found that as soon as the air purifier was turned on, 83.4% of S. epidermidis, 81.4% of B. subtilis, 88.5% of A. niger, and 75.8% of P. citrinum were removed from the air. In a hospital environment, the PCO air purifier efficacy was lower than that in the chamber. Besides, relative humidity, distances from the air purifier and room size were suspected to affect the efficacy of the photocatalytic filters.