To evaluate the repeatability of corneal biomechanical parameters in normal and keratoconus eyes, and explore factors that affects the repeatability, and further assess the diagnostic ability of new parameters. Seventy-seven keratoconus eyes of 47 patients and 77 right eyes of 77 normal subjects were recruited in current study. All participants received three repeated measurements with 2 to 5 minutes interval. The interclass correlation coefficient (ICC), Cronbach' α and repeatability coefficient (RC) were evaluated. The liner regression analysis was used to identify factors that affect the repeatability, and linear mixed effects model was performed to compare the parameters differences. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic ability of new parameters. Eighteen parameters in normal eyes and twenty-two parameters in keratoconus eyes showed excellent repeatability (ICC ≥ 0.90). Age, axial measurement (AL), spherical equivalent, astigmatism, gender, mean keratometry (Kmean), intraocular pressure (IOP) and central corneal thickness (CCT) could affect the repeatability of new Corvis ST parameters. Compared with normal eyes, the Ambrósio's Relational Thickness horizontal (ARTh), biomechanical corrected IOP (bIOP), stiffness parameter at first applanation (SP A1) were low and the Max Inverse Radius, deformation amplitude (DA) Ratio Max [2 mm], Pachy Slope, DA Ratio Max [1 mm], Integrated Radius and Corvis Biomechanical Index (CBI) were high in keratoconus eyes (All P < 0.05). Both ARTh and CBI had high Youden index (0.870), and the corresponding cut-off values were 379.29 and 0.44. The repeatability of Corvis ST parameters was acceptable both in normal and keratoconus eyes, and new parameters could effectively diagnose keratoconus eyes from normal eyes.