Objective To explore the diagnostic performance of 64-slice spiral CT combined with exercise treadmill test for coronary artery disease (CAD). Methods Eighty-six patients suspected of CAD were divided into low risk group, intermediate risk group and high risk group according to estimated pretest probabilities of CAD. All patients underwent coronary angiography, 64-slice spiral CT and exercise treadmill test. With coronary artery stenosis rate >50% as positive findings, the sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of 64-slice spiral CT and 64-slice spiral CT combined with exercise treadmill test in diagnosis of CAD were calculated. Results With coronary angiography as the "golden criteria", the sensitivity, specificity, PPV, NPV and accuracy of 64-slice spiral CT in diagnosis of CAD were 95.2%, 88.6%, 88.9%, 95.1% and 91.9%, respectively. The sensitivity of low risk group, intermediate risk group and high risk group was 100%, 100% and 92.6%, specificity was 94.4%, 94.1% and 66.7%, PPV was 80.0%, 91.7% and 89.3%, NPV was 100%, 100% and 75.0%, and accuracy was 95.5%, 96.4% and 86.1%, respectively. The sensitivity, specificity, PPV, NPV and accuracy of 64-slice spiral CT combined with exercise treadmill test in diagnosis of CAD were 97.6%, 97.7%, 97.6%, 97.7% and 97.7%, respectively. Conclusion 64-slice spiral CT combined with exercise treadmill test works well in screening CAD, especially for those with a low or intermediate estimated pretest probability.