We evaluated the results of sequential SPECT dual-isotope imaging with Tl-201 and Tc-99m MIBI in 24 patients, all of whom also had coronary angiography within the past one month. Coronary angiography showed that 12 patients had no CAD, 4 patients had one-vessel CAD, 7 patients had two-vessel CAD and 1 patient had three-vessel CAD. Serial studies of resting Tl-201 and dipyridamole stress Tc-99m MIBI were completed within 2 hours. When more then 50% of coronary artery narrowing was considered significant. The overall sensitivity and specification of CAD detection were 91.7%. The sensitivity of CAD detection in patients with one-vessel and multi-vessel diseases was 75% and 100%, respectively. Therefore, sequential dual-isotope SPECT demonstrated high sensitivity and specificity of CAD detection. In conclusion, sequential dual-isotope imaging is feasible and can be completed in a short time and may therefore enhance laboratory throughput and patient convenience.