The aim of this study was to validate a motion-immune (MI) solution to dynamic CT myocardial perfusion measurement, in the presence of motion without image registration. The MI perfusion technique was retrospectively validated in six swine (37.3 ± 7.5 kg) with a motion-susceptible (MS) perfusion technique performed for comparison. In each swine, varying severities of stenoses were generated in the left anterior descending (LAD) coronary artery using a balloon under intracoronary adenosine stress, followed by contrast-enhanced imaging with 20 consecutive volume scans per stenosis. Two volume scans were then systematically selected from each acquisition for both MI and MS perfusion measurement, where the resulting LAD and left circumflex (LCx) measurements were compared to reference microsphere perfusion measurements using regression and diagnostic performance analysis. The MI (PMI) and microsphere (PMICRO) perfusion measurements were related through regression by PMI = 0.98 PMICRO + 0.03 (r = 0.97), while the MS (PMS) and microsphere (PMICRO) perfusion measurements were related by PMS = 0.62 PMICRO + 0.15 (r = 0.89). The accuracy of the MI and MS techniques in detecting functionally significant stenosis was 93% and 84%, respectively. The motion-immune (MI) perfusion technique provides accurate myocardial perfusion measurement in the presence of motion without image registration.