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Accurate assessment of global and regional right ventricular (RV) systolic function is challenging. The aims of this study were to confirm the reliability and feasibility of a three-dimensional (3D) speckle-tracking echocardiography (STE) system, using comparison with cardiac magnetic resonance imaging (CMR), and to assess the contribution of regional RV function to global function.In a retrospective, cross-sectional study setting, RV volumetric data were studied in 106 patients who were referred for both CMR and 3D echocardiography within 1 month. Three-dimensional STE-derived area strain, longitudinal strain, and circumferential strain were assessed as global, inlet, outflow, apical, and septal segments.Seventy-five patients (70%) had adequate 3D echocardiographic data. RV measurements derived from 3D STE and CMR were closely related (RV end-diastolic volume, R2 = 0.84; RV end-systolic volume, R2 = 0.83; RV ejection fraction [RVEF], R2 = 0.70; P < .001 for all). RVEF and RV end-diastolic volume from 3D STE were slightly but significantly smaller than CMR values (mean differences, −2% and −10 mL for RVEF and RV end-diastolic volume, respectively). Among conventional echocardiographic parameters for RV function (tricuspid annular plane systolic excursion, fractional area change, S′ of the tricuspid annulus, RV free wall two-dimensional longitudinal strain), only fractional area change was significantly related to RVEF (r = 0.34, P = .003). Among segmental 3D strain variables, inlet area strain (r = −0.56, P < .001) and outflow circumferential strain (r = −0.42, P < .001) were independent factors associated with CMR-derived RVEF.RV volume and RVEF determined by 3D STE were comparable with CMR measurements. Regional RV wall motion showed that heterogeneous segmental deformations affect global RV function differently; specifically, inlet area strain and outflow circumferential strain were significant factors associated with RVEF in patients with underlying heart diseases.We present a novel 3D speckle-tracking echocardiographic measurement system for the assessment of right ventricular systolic function.This system provides a reliable measurement of right ventricular systolic function using an echocardiographic methodology.The right ventricular regional strain derived by this system was revealed to contribute differently to the right ventricular ejection fraction.