ECG-gated dual-isotope acquisition protocol involving rest imaging with 201Tl and stress 99mTc-tetrofosmin (TF) SPECT was designed for the simultaneous assessment of rest/stress myocardial perfusion and rest ventricular systolic function. This study assessed the feasibility and diagnostic accuracy of this protocol. Forty-five patients underwent the dual-isotope SPECT protocol. Twenty minutes after resting injection of 111 MBq of 201Tl, 370 MBq of 99mTc-TF was administered at a peak exercise. The dual-isotope gated SPECT acquisition was performed 1 hour later. Then, the regional count increase rate (%WT) of 99mTc-TF from end-diastole end-systole was calculated using an automated method which was developed for quantification of regional wall thickening based on circumferential profile analysis in our laboratory. Myocardial perfusion and contractility analysis was carried out using 8 segments of left ventricle with comparison of coronary angiographical findings. The sensitivity and specificity for the detection of diseased coronary vessels (> = or 75% stenosis) were 76% and 94%, respectively. Infarcted regions showing reversible defect had significantly greater %WT as compared with those with fixed defects (63 seg; 12.7 +/- 6.1% vs. 36 seg; 8.9 +/- 7.2%, p < 0.01). In conclusion, this dual-isotope protocol has some advantages; i.e., shortening an examination time, having the exact registration of stress/rest perfusion, and simultaneous evaluation of resting regional wall thickening.