Myocardial perfusion imaging with dipyridamole is an accepted method for diagnosing coronary artery disease. However, the simultaneous effect of dipyridamole on perfusion and function in the detection of coronary artery disease has not been studied extensively. The aim of this study was to investigate this effect using technetium-99m methoxy isobutyl isonitrile (Tc-MIBI) imaging. Twenty-eight patients with proven coronary artery disease participated in a 2-day protocol (rest and then 0.56 mg/kg dipyridamole i.v.) using Tc-MIBI for first-pass radionuclide ventriculography followed by SPECT imaging. Fifteen patients (54%) demonstrated a decrease in left ventricular ejection fraction from rest to dipyridamole and/or wall motion abnormality, while 21 patients (75%) showed abnormal perfusion by dipyridamole MIBI SPECT imaging. Concordance between perfusion and function was 65%. Correlation for one-, two- and three-vessel disease (coronary angiography) to perfusion versus function were: 54% vs. 38%, 80% vs. 60% and 80% vs. 80%, respectively. Left ventricular dysfunction using dipyridamole was noted in patients with multivessel disease, and with multi-reversible perfusion filling defects. We conclude that dipyridamole in combination with Tc-MIBI for assessment of perfusion and function serves as a valuable tool to identify patients with multivessel disease and a high amount of myocardium at risk.