Current guidelines recommend routine second transurethral resection (TUR) for accurate diagnosis and to prevent understaging of muscle-invasive bladder cancer. We evaluated the diagnostic accuracy of immediate second resection of the tumor bed during initial TUR and its prognostic significance. Patients (n=126) undergoing TUR were prospectively randomized to undergo (n=63) or not undergo (n=63) immediate second resection of the tumor bed after complete TUR. Second resection was repeated until muscularis propria (MP) was identified in the specimen and the depth of tumor invasion was inspected. The results of second resection were compared with final pathology results for diagnostic accuracy. Recurrence and progression rates were compared in the two groups, and factors affecting recurrence were evaluated. Patient age, sex distribution, number of tumors, pathologic T stage and grade were similar in the groups. MP was included in all TUR specimens in the immediate second resection group, compared with 41 of 63 (65.1%) in the nonsecond resection group. The concordance rate of second resection with final pathology was r=0.810 (P<0.01). The sensitivity and specificity of second resection for T(2) disease were 90.9% and 98.0%, respectively, and the positive and negative predictive values of second resection for T(2) disease were 90.0% and 96.2%, respectively. Among the 94 patients followed up, those in the second resection group had significantly higher 2-year recurrence-free survival rate (77.0% vs 45.8%, P=0.025), but there was no difference in progression-free survival rate. Immediate second resection of the tumor bed after complete TUR improves the effectiveness of resection by immediately confirming the presence of MP in the specimen and accurately differentiating muscle-invasive disease. The advantages of immediate second resection were precise prediction of final pathology results and reduced early recurrence.