Objectives: The study aimed to evaluate the diagnostic performance of optical coherence tomography (OCT) in identifying functionally significant coronary stenosis in a vessel with single stenosis. Background: The OCT-based morphofunctional computational method for deriving the optical flow ratio (OFR) has diagnostic value, as it can identify the functional severity of coronary stenosis, but the ability of the OFR to aid the OCT in determining coronary stenosis hemodynamics in single-stenosis lesion remains unclear. Methods: 74 vessels with single stenosis were studied in 69 patients; all cases were performed through OCT and quantitative flow ratio (QFR), and OCT images were used to perform OFR. Results: Among vessels with single stenosis, OFR showed a good correlation with QFR (r = 0.86; p < 0.001). Taking QFR as the standard, the vessel-level diagnosis accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of OFR were 90% (95% CI: 81 to 96), 94% (95% CI: 77 to 99), 88% (95% CI: 74 to 96), 85% (95% CI: 68 to 94) and 95% (95% CI: 82 to 99), respectively. Among vessels with OFR/QFR concordance, both the minimum lumen area (MLA) and minimum lumen diameter (MLD) showed excellent diagnostic efficiency (MLA: area under the curve (AUC) = 0.92, 95% CI: 0.85 to 0.98, p < 0.001; MLD: AUC = 0.93, 95% CI: 0.86 to 0.98, p < 0.001) in determining the functional significance of coronary stenosis in a single stenosis lesion, and the best cutoff values were 1.55 mm2 and 1.40 mm. Conclusions: OFR has a good correlation with QFR. OCT-measured MLA and MLD have excellent diagnostic efficiency in identifying the hemodynamic significance of coronary stenosis in a vessel with single stenosis.