Myocardial perfusion imaging (MPI) by gated single-photon emission computed tomography (SPECT) is a feasible method in the evaluation of left ventricular perfusion and function. The purpose of this study was to determine the threshold and grading of left ventricular (LV) diastolic dysfunction (LVDD) using gated SPECT MPI. A total of 149 patients were recruited in the study. All of the patients underwent a standard 2-day stress/rest gated MPI study and transthoracic echocardiography within 2 weeks. The reconstructed rest-only images were analyzed by Cedar-Sinai's quantitative gated SPECT and the LV diastolic parameters, including peak filling rate (PFR), time to PFR (TTPF) and secondary PFR (PFR2) to PFR ratio were provided and compared to echocardiographic data. 68 (45.6%) and 81 (54.4%) of patients were categorized in LVDD-absent and LVDD-present groups on the basis of LVDD evidence in echocardiography, respectively. receiver-operating-characteristic analysis for PFR and TTPF was performed, resulting in diagnostic sensitivities of 70 and 57% and specificities of 60 and 75% for PFR <2.6 end-diastolic volumes (EDV)/s and TTPF>160.5 ms, respectively. Applying our previously used thresholds of <1.70 EDV/s for PFR, >208 ms for TTPF and >1 for PFR2/PFR, sensitivities and specificities of 9.9 and 96.6%, 9.9 and 95.6% and 13.8 and 88% were resulted, respectively. Grading of LVDD on the basis of MPI-obtained diastolic parameters showed considerable overlapping data by interquartile range. Gated SPECT MPI can be used as a highly specific means for detection of LV diastolic dysfunction when compared to echocardiography. However, grading of severity of diastolic heart failure appears to be impracticable.