BACKGROUND The value of layer-specific two-dimensional speckle tracking echocardiography (LS2D-STE) for evaluating viable myocardium (VM) in patients with acute myocardial infarction (AMI) was unclear, this study provides new insights into it and to make a comparison with dualisotope simultaneous acquisition single photon emission computed tomography ( DISA-SPECT). METHODS Forty hospitalized patients with AMI and left ventricular systolic dysfunction (left ventricular ejection fraction <50%) underwent LS2D-STE and DISA-SPECT before percutaneous coronary intervention (PCI). The longitudinal, circumferential, and radial peak systolic strains and the peak systolic strain rates of 3 myocardiallayers (endocardium, mid-myocardium, and epicardium), as well as the total wall thickness, were determined by LS2D-STE. Routine echocardiography was followedup at 1, 3, 6 months after PCI, with the improvement of the wall motion as the goldenstandard for evaluating VM. RESULTS The sensitivity, specificity and accuracy of DISA-SPECT for evaluating VM were 82.1%, 74.3%, and 79.3%, respectively. Among the layer-specific parameters, only endocardial (endo-) longitudinal strain (LS) and endo- longitudinal strain rate (LSr) were used as independent parameters for evaluating VM (P < .05), and the sensitivity, specificity and accuracy of endo-LS and endo-LSr in evaluation of VM were 77.1%, 65.4%, and 72.9% vs 72.9%, 65.4%, and 69.7%. Endo-LS and endo-LSr were superior to total wall thickness LS and LSr (AUC endo-LS 0.767 vs total-LS 0.669; endo-LSr 0.743 vs total-LSr 0.682). The parallel test and the serial test of combination of endo-LS and endo-LSr showed similar sensitivity, specificity and accuracy to DISA-SPECT (P > .05). CONCLUSIONS The endo-LS and endo-LSr analysis of LS2D-STE can evaluate the VM well, and its sensitivity, specificity and accuracy in detection of VM are similar to those of DISA-SPECT, resulting in LS2D-STE being a good option for the assessment of VM.