The aim of this study was to assess Corvis ST biomechanical indices in reference to corneal enantiomorphism. In a prospective observational cohort study, 117 eyes from 63 patients with normal or keratoconus corneas were assessed by three independent observers. In the control group (n = 62), no significant differences were observed between the three observers for all indices. The best reproducibility was obtained with pachymetry and the weakest with CBI. All indices but CBI and arc length featured COV < 10%. All indices except the PD and SSI correlated with pachymetry; all but Rad correlated with IOP. The comparison of the thinnest with the thickest corneas showed no significant differences for any index except pachymetry. In the keratoconus group (n = 55), loss of corneal enantiomorphism was confirmed for all indices except the arc length, velocity, and PD. Significant differences between both groups were found for all indices, even after adjustment for pachymetry and intraocular pressure. The CBI featured the best accuracy (92%), sensitivity (91%), and graphical relevance for keratoconus diagnosis. However, its reproducibility was weak in normal corneas and was strongly dependent on corneal thickness. The SSI was independent of corneal thickness, highly reproducible, and provided the expected enantiomorphism characteristics in both groups, making it a relevant biomarker of biomechanical corneal behavior.