Myocardial perfusion defect, assessed with single photon emission computed tomography (SPECT), is useful for patient management and risk stratification. Left ventricle Global Longitudinal Strain (LV GLS) has gained interest for observing subclinical LV dysfunction. We aimed to investigate the utility of LV GLS in evaluating myocardial perfusion defect. A retrospective study of all patients who underwent SPECT and LV GLS at Tel Aviv Sourasky medical center. Overall, 86 patients were included. LV GLS and SPECT correlated in the base and apex sections for infraction, and in the apex only for ischemia. Adjusted analysis showed a significant correlation between LV GLS of both the mid and apical section and infarction by SPECT, but no association with ischemia. No associations were found by arterial supply territory. A sub-analysis of patients without left bundle branch block (LBBB) strengthened the correlations, with a 58–70% higher chance of both fixed and reversible defects for every 1-unit decrease LV GLS in the mid and apical sections. LV GLS effectively evaluated the presence of infarction by SPECT in the mid and apical sections, particularly in patients without LBBB. Due to its high availability, LV GLS may have a role in evaluating myocardial perfusion defect.