Neural selectivity of orientation is a fundamental property of visual system. We aim to investigate whether and how the orientation selectivity changes in amblyopia.Seventeen patients with amblyopia (27.1 ± 7.1 years) and 18 healthy participants (25.1 ± 2.7 years) took part in this study. They were asked to continuously detect vertical gratings embedded in a stream of randomly oriented gratings. Using a technique of subspace reverse correlation, the orientation-time perceptive field (PF) for the atypical grating detection task was derived for each participant. Detailed comparisons were made between the PFs measured with the amblyopic and healthy eyes.The PF of the amblyopic eyes showed significant differences in orientation and time domain compared with that of the normal eyes (cluster-based permutation test, ps < 0.05), with broader bandwidth of orientation tuning (31.41 ± 10.59 degrees [mean ± SD] vs. 24.76 ± 6.85 degrees, P = 0.039) and delayed temporal dynamics (483 ± 68 ms vs. 425 ± 58 ms, P = 0.015). None of the altered PF properties correlated with the contrast sensitivity at 1 cycle per degree (c/deg) in amblyopia. No difference in PFs between the dominant and non-dominant eyes in the healthy group was found.The altered orientation-time PF to the low spatial frequency and high contrast stimuli suggests amblyopes had coarser orientation selectivity and prolonged reaction time. The broader orientation tuning probably reflects the abnormal lateral interaction in the primary visual cortex, whereas the temporal delay might indicate a high level deficit.