Background: Real-time (RT) myocardial contrast echocardiography (MCE) is a novel method for the assessment of regional myocardial perfusion. We sought to evaluate the feasibility and diagnostic accuracy of quantitative RT-MCE in predicting significant coronary stenosis, with reference to quantitative coronary angiography. Hypothesis: RT-MCE can identify anatomically significant coronary artery stenosis in selected patients. RT-MCE is probably an effective method for detection of angiographically significant coronary artery stenosis. Methods: Thirty-five patients (mean age, 59.94 ± 10.63 years; 25 males) scheduled for coronary angiography underwent RT-MCE at rest, and shortly afterward underwent gated single-photon emission computed tomography (gated-SPECT). Coronary angiography was performed within 1 week after RT-MCE in all patients. The observing indexes included the images of RT-MCE that were analyzed quantitatively from microbubble replenishment curves for myocardial perfusion by using the Q-Lab software. The sensitivity and specificity of RT-MCE for quantitative detection of coronary artery disease (CAD) were obtained. The receiver operator characteristic (ROC) curves were used to assess the differences of accuracy in ischemic segments with A, β and A × β respectively. The sensitivity and specificity of gated-SPECT and RT-MCE for assessment of CAD were calculated using a 4-score method. Results: A total of 513 segments among 595 segments in 35 patients were obtained. The cutoffs for A, β and A × β were 4.58, 0.64, and 2.73, and the sensitivity and specificity of quantitative RT-MCE for detection of CAD were 86.0%, 80.2%, 88.9%, and 84.1%, 64.6%, 79.9%, respectively. Meanwhile, the sensitivity and specificity of semiquantitative analysis for assessment of CAD were 66.7% and 61.8%. The ROC curve area of A and A × β was 0.91 and 0.90 in the middle segments. The ROC area of A was 0.52 in the base segments. The sensitivity and specificity of gated-SPECT for assessment of CAD were 84.8% and 82.7%, respectively. The sensitivity of multi-indexes RT-MCE increased. The sensitivity was 89.1%, 90.4%, and 96.3% by A + β, A + A × β, and β + A × β. Conclusions: Quantitative RT-MCE is an effective method for the detection of coronary artery stenosis. Quantitative RT-MCE is segmented for assessment to ischemic myocardium. RT-MCE with multi-indexes has a valuable application for assessment of CAD surpassing SPECT.