As controlling temperature and humidity is crucial for maintaining comfort and preventing microbial growth, operating rooms (ORs) are the most energy-intensive areas in hospitals. We aimed to evaluate the energy consumption of three dehumidification air conditioning systems used in ORs and their corresponding air quality for ORs at rest. This study selected three ORs using a conventional heating, ventilation, and air conditioning (HVAC) system; a liquid desiccant air conditioning (LDAC) system; and a rotary desiccant air conditioning (RDAC) system, respectively. The indoor thermal–hygrometric conditions, air quality, and energy consumption of the ORs were monitored in this study. The median levels of relative humidity (RH) were 66.7% in the OR using the conventional HVAC system, 60.8% in the OR using the LDAC system, and 60.5% in the OR using the RDAC system. The median daily total energy consumption of the RDAC system (10.1 kWh/m²) and LDAC system (11.8 kWh/m²) were 28.12% and 16.54% lower, respectively, than that of the conventional HVAC system (14.1 kWh/m²). The PM_≥0.5 levels and airborne bacterial concentrations in the ORs met the ISO 14644-1 Class 7 standard and China’s GB50333-2013 standard, respectively. The RDAC system was clearly superior to the LDAC and conventional HVAC systems in terms of energy consumption.