A negative pressure isolation ward prevents the outflow of airborne microorganisms from inside the ward, minimizing the spread of airborne contamination causing respiratory infection. In response to recent outbreaks of severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), Korea has increased the number of these facilities. However, airborne contaminants that flow into the ward from adjacent areas may cause secondary harm to patients. In this study, the sterilization effect of upper-room ultraviolet germicidal irradiation (UR-UVGI) on microorganisms generated within the negative pressure isolation ward and those flowing inward from adjacent areas was evaluated through field experiments and computational fluid dynamics (CFD) analysis, to assess the potential of this approach as a supplementary measure to control such microorganisms. The sterilization effect was found to be not high because of high-level ventilation. CFD analysis under various conditions shows that the sterilization effect for indoor-generated microorganisms varies with the level of UV radiation, the source locations of the indoor-generated microorganisms, air supplies and exhausts, the UVGI system, and the airflow formed under the specified conditions. Our results show that when the UVGI system is installed in the upper part of the ward entrance, contaminated air from adjacent area is strongly sterilized.