Objective To detect circulating tumor cells (CTCs) in peripheral blood from the patients with bladder cancer using subtraction enrichment and immunostaining-fluorescence in situ hybridization (SE-iFISH) and to explore the relation between CTCs detection result and the clinicpathological parameters of tumors. Methods Sixteen patients 〔(66.88±8.97) yr.〕 with bladder cancerwere included, whlile 10 normal people 〔(61.90±9.13) yr.〕 were enrolled as control. Peripheral blood was harvested (6 mL per case) for the detection of CTCs with the method of subtraction enrichment and SE-iFISH, and the correlations of CTCs level to the clinicpathological parameters of tumors were analyzed, including hematuria, size, number, NMIBC or MIBC, stage, grade and pathological type. Results Positive rates of CTCs in experimental and control group were 43.75% (7/16) and 0% (0/10), respectively. The difference was statistically significant (P=0.022 7). The number of CTCS was not associated with the clinicpathological parameters of bladder cancer, which probably due to small sample size. Conclusion It is applicable to detect CTCs of bladder cancer by SE-iFISH.