Amblyopia screening during childhood is critical for early detection and successful treatment. In the current study, we develop and evaluate a screening method that exploits the imbalanced interocular inhibition between amblyopic and fellow eyes. In nineteen subjects with anisometropic amblyopia and twenty-two age-matched subjects with myopia, we measured the area under the contrast sensitivity functions (AUCSFs) in eight monocular conditions defined by the tested eye (left, right), patching of the untested eye (translucent, opaque), and refractive status (corrected, uncorrected). For each tested eye, we defined the inhibition index as the ratio between the AUCSF values obtained in the translucent and opaque patching conditions of the untested eye. To evaluate the screening potential of the inhibition index, we compared results from patients with amblyopia and myopia. With and without optical correction, the index was significantly lower in the amblyopic eye than in the fellow eye of the amblyopic subjects and both eyes of the myopic subjects. No significant difference was found among the two eyes of the myopic subjects and the fellow eyes of the amblyopic subjects. With the inhibition index as the predictor, a logistic regression model successfully discriminated amblyopic eyes from myopic eyes with 100% accuracy in the uncorrected condition. In the corrected condition, with the inhibition index and interocular visual acuity difference as predictors, amblyopic eyes were likewise discriminated from myopic eyes with 100% accuracy. This pattern of CSF changes, caused by the different patching modes of the untested eye, provides a potential CSF signature to discriminate anisometropic amblyopia from myopia.