Objective To assess the value of two-dimentional strain/strain rate (2D-S/SR) imaging compared with endomyocardial biopsy (EMB) in detecting acute rejection (AR) reaction after heart transplantation (HT). Methods Twenty-five patients who required HT underwent echocardiography within 12 h after EMB. Ten patients of grade 0 AR were regarded as group A, 8 patients with grades Ⅰa-Ⅰb were considered as group B, 7 patients with grades ≥Ⅱa were considered as group C. Thirty age-matched normal controls were considered as group D. The longitudinal, radial and circumferential stain and strain rate of different ventricular wall in systolic were measured and compared among groups. Results Compared with the group D, longitudinal stain (LS) of basal-septum, circumferential strain rate (CSR) of ant-septum at the level of mitral valve, radial strain (RS) of all segments at the level of mitral valve reduced in groups A, B and C. Conclusion The RS of 2D-S/SR is an effective and sensitive technique for the detection of acute rejection, and longitudinal stain can be used as an early index of AR for grade ≥Ⅱ after HT.