In conventional coronary computed tomography angiography (CCTA), metal artifacts are frequently observed where stents are located, making it difficult to evaluate in-stent restenosis. This study was conducted to investigate whether subtraction CCTA can improve diagnostic accuracy in the evaluation of in-stent restenosis. Subtraction CCTA was performed using 320-row CT in 398 patients with previously placed stents who were able to hold their breath for 25 s and in whom mid-diastolic prospective one-beat scanning was possible. Among these patients, 126 patients (94 men and 32 women, age 74 ± 8 years) with 370 stents who also underwent invasive coronary angiography (ICA) were selected as the subjects of this study. With ICA findings considered the gold standard, conventional CCTA was compared against subtraction CCTA to determine whether subtraction can improve diagnostic accuracy in the evaluation of in-stent restenosis. When non-assessable stents were considered to be stenotic, the diagnostic accuracy in the evaluation of in-stent restenosis was 62.7 % for conventional CCTA and 89.5 % for subtraction CCTA. When the non-assessable stents were considered to be non-stenotic the diagnostic accuracy was 90.3 % for conventional CCTA and 94.31 % for subtraction CCTA. When subtraction CCTA was used to evaluate only the 138 stents that were judged to be non-assessable by conventional CCTA, 116 of these stents were judged to be assessable, and the findings for 109 of them agreed with those obtained by ICA. Even for stents with an internal diameter of 2.5-3 mm, the lumen can be evaluated in more than 80 % of patients. Subtraction CCTA provides significantly higher diagnostic accuracy than conventional CCTA in the evaluation of in-stent restenosis.