Purpose: Increased incidence of anterior cruciate ligament injuries has amplified the need for quantitative research in clinical and academic settings. We used a novel digital arthrometer to measure knee laxity in healthy people and patients with anterior cruciate ligament injuries. Changes in stiffness were also assessed to develop new indicators for detecting anterior cruciate ligament injury. The purpose of this study was to use arthrometer to measure the quantitative indicator of knee laxity, bringing clinicians a new perspective on how to identify injury to the ACL.Methods: In this cross-sectional study, anterior tibial displacement under continuous loading was measured using a novel digital arthrometer in 30 patients with unilateral anterior cruciate ligament injury and 30 healthy controls. Load-displacement curves were plotted, using real-time load and displacement changes. Stiffness was defined by the slope of the applied load to tibial displacement. Anterior tibial displacement and instantaneous stiffness values under different loads were compared. The restricting contribution of the anterior cruciate ligament transformed the displacement-stiffness curve from a sharp decrease to a stable increase, resulting in a minimum stiffness value. Using the minimum stiffness as the turning point, the load-displacement curve was divided into regions 1 and 2. The two regions’ stiffness changes were compared. Based on the findings, receiver operating characteristic curves were plotted and the area under the curve was calculated to estimate the diagnostic accuracy.Results: Anterior tibial displacement was significantly greater in the anterior cruciate ligament injury group than in the controls under each 10-N increase load (p < 0.05). In the anterior cruciate ligament injury group, instantaneous stiffness was significantly lower on the injured side than on the healthy side (p < 0.05). In the two regions of the load-displacement curve, stiffness was significantly lower in the anterior cruciate ligament injury group than in the control group (all, p < 0.05). Receiver operating characteristic curves were plotted, using changes in stiffness under the two regions in both groups. Stiffness in region 2 had the largest area under the curve (0.94; 95% CI, 0.88–0.99). Using the cut-off value of 9.62 N/mm to detect ACL injury, the sensitivity and specificity were 93% and 82%, respectively.Conclusion: Our investigation of ligament stiffness provides novel insights into the properties of knee laxity. Stiffness in the later stages of increased loading <9.62 N/mm could be a valid indicator for identifying knee laxity.