Rapid and efficient detection of acetone gas at room temperature is of great importance for non-invasive screening of diseases as well as environmental and industrial safety. In this work, we have demonstrated a resistive room temperature acetone gas sensor based on molybdenum disulfide (MoS2)-copper oxide (CuO) nanocomposites. The morphological and structural characteristics, crystalline nature, and attachment of CuO nanoparticles on MoS2 nanosheets were investigated using various standard material characterization techniques. The sensing properties of the MoS2-CuO nanocomposite sensor were investigated systematically by exposing to various acetone gas concentrations. It was found that the sensor showed excellent response towards acetone with high sensitivity and fast response and recovery time with a detection limit of 93 ppb. Furthermore, the sensor also showed good repeatability, reproducibility and stability, and portended enormous potential for acetone gas monitoring in the fields of health care, environmental, and industrial applications. The probable sensing mechanism of the MoS2-CuO nanocomposite based acetone sensor has been discussed in detail.