A sensing module including a sample loading layer, a sensing layer and an optical resonance layer locating between the sample loading layer and the sensing layer is provided. The sample loading layer includes at least a sample loading depression, and the sample loading depression exposes part of the optical resonance layer, and the sample loading depression is adapted to load sample. A surface of the optical resonance layer has optical resonance structures, and the optical resonance structures are located beside bottom of the sample loading depression or below the bottom of the sample loading depression. The sensing layer is configured to receive light and turn it into electrical signals. A sensing method is also provided.