Purpose: N-13 ammonia uptake and retention in the myocardium is related to perfusion and metabolism. There are several potential advantages of N-13 ammonia positron emission tomography (PET) to detect myocardial ischemia, such as higher spatial resolution, greater counting efficiencies, and robust attenuation correction. But there are few reports comparing Tc-99m myocardial perfusion single photon emission tomography (MPS) and N-13 ammonia PET. We thus compared adenosine stress N-13 ammonia PET/CT and Tc-99m sestamibi MPS in patients with suspected coronary artery stenosis. Materials and Methods: Seventeen patients (male 13 : 63¡¾11 years old) underwent adenosine stress N-13 ammonia PET/CT (Discovery ST, GE), Tc-99m sestamibi MPS (dual head gamma camera, Hawkeye, GE) and coronary angiography within 1 week. N-13 ammonia PET/CT and Tc-99m sestamibi MPS images were assessed with a 20-segment model by visual interpretation and quantitative analysis using automatic quantitative software (Myovation, GE). Results: Both sensitivities and specificities of detecting an individual coronary artery stenosis were higher for N-13 ammonia PET/CT than Tc-99m sestamibi MPS (PET/CT: 91%/ 89% vs MPS: 65%/ 82%). N-13 ammonia PET/CT showed reversibility in 52% of segments that were considered non-reversibile by Tc-99m sestamibi MPS. In the 110 myocardial segments supplied by the stenotic coronary artery, N-13 ammonia PET/CT showed higher count densities than Tc-99m MPS on rest study (p < 0.01), and the difference of count density between the stress and the rest studies was also larger on N-13 ammonia PET/CT. Conclusion: Adenosine stress N-13 ammonia PET/CT had higher diagnostic sensitivity and specificity, more reversibility of perfusion defects and greater stress/rest uptake differences than Tc-99m sestamibi MPS. Accordingly, N-13 ammonia PET/CT might offer better assessment of myocardial ischemia and viability. (Nucl Med Mol Imaging 2008;42(5):354-361)