Ammonia gas-sensing properties of the iodinedoped polythiophene film deposited by electrophoretic deposition technique on an epoxy glass substrate with concentric circular electrodes were developed. The gas-sensing properties were investigated for different concentrations of ammonia gas from 0.046 to 0.185 vol% at room temperature. The obtained results demonstrate that the prepared film has a high sensitivity and excellent repeatability as well as a great potential for ammonia gas-sensing application at room temperature. Furthermore, ammonia gas-sensing mechanism of the resistance-based sensor is discussed.