To investigate the diagnostic value and reproducibility of deformable registration algorithm (DRA)-derived mechanical dyssynchrony parameters in dilated cardiomyopathy (DCM) patients. The present study included 80 DCM patients (40 with normal QRS duration (NQRS-DCM); 40 with left bundle branch block (LBBB-DCM)) and 20 healthy volunteers. The balanced steady-state free-precession (bSSFP) cine images were acquired using a 3.0T scanner. Mechanical dyssynchrony parameters were calculated based on DRA-derived segmental strain, including uniformity ratio estimate (URE) and standard derivation of time-to-peak (T2Psd) parameters in circumferential, radial, and longitudinal orientations. DCM patients showed significant mechanical dyssynchrony reflected by both URE and T2Psd parameters compared with controls. Among DCM patients, LBBB-DCM showed decreased CURE (0.78 ± 0.21 vs. 0.93 ± 0.05, p < 0.001) and RURE (0.69 ± 0.14 vs. 0.83 ± 0.15, p = 0.001), and increased T2Psd-Ecc (median with interquartile range, 94.1 (54.4-123.2) ms vs. 63.7 (44.9-80.4) ms, p = 0.003) and T2Psd-Err (91.1 (61.1-103.2) ms vs. 62.3 (46.3-104.5) ms, p = 0.041) compared with NQRS-DCM patients. CURE showed a strong correlation with QRS duration (r = - 0.54, p < 0.001), with maximum AUC (0.791) to differentiate LBBB-DCM from NQRS-DCM patients. Improved intra- and inter-observer reproducibility was found using URE indices (coefficient of variation (CoV), 1.20-3.17%) than T2Psd parameters (CoV, 15.28-41.18%). The DRA-based CURE showed significant correlation with QRS duration and the highest discriminatory value between LBBB-DCM and NQRS-DCM patients. URE indices showed greater reproducibility compared with T2Psd parameters for assessing myocardial dyssynchrony in DCM patients. • The strain analyses based on DRA suggested that DCM patients have varying degrees of mechanical dyssynchrony and there is a significant difference from normal controls. • CURE showed the strongest correlation with QRS duration and was the best parameter for differentiating DCM patients with normal QRS duration from patients with LBBB, and with normal controls. • URE indices showed improved reproducibility compared with T2Psd parameters in all three orientations (circumferential, radial, and longitudinal).