Objective To explore the diagnostic value of urinary exosomes nano-enrichment chips combined with pathological markers for the early diagnosis of bladder cancer. Methods A total of 110 patients suspected of having bladder cancer who were treated at the Fifth Affiliated Hospital of Zhengzhou University and 110 volunteers who underwent health check-ups at the same time were prospectively included as subjects. They were randomly divided into the chip detection group (n＝55) and the conventional detection group (n＝55). Based on the results of pathological biopsy, the patients were divided into the bladder cancer group (n＝58) and the nonbladder cancer group (n＝52). The chip detection group used nano-chips to enrich urinary exosomes, while the conventional detection group used ultracentrifugation to enrich urinary exosomes. The differences in extraction time, recovery rate and protein purity of the exosomes between the two groups were compared. The expression differences of microRNA-21 and miR-200b were detected by real-time quantitative polymerase chain reaction (RT-qPCR); the diagnostic efficacy of single and combined pathological markers for early bladder cancer was evaluated by the receiver operator characteristic (ROC) curve. Results The extraction time, recovery rate and protein purity of exosomes using the nano-chip method were significantly better than those of the ultracentrifugation method (all P＜0.001). Compared with the non-bladder cancer group, the expression of miR-21 and miR-200b in the exosomes of the bladder cancer group was significantly upregulated (all P＜0.001). The receiver operator characteristic (ROC) curve analysis indicated that the area under the curve (AUC) of the combined detection of the two pathological markers reached 0.910, with a sensitivity of 89.1% and a specificity of 86.7%. This was significantly superior to either single indicator (all P＜0.05). Conclusion The combined use of urinary exosomes nano-enrichment chips and miR-21 and miR-200b detection can significantly enhance the diagnostic efficacy of early-stage bladder cancer, breaking through the limitations of the insufficient accuracy of traditional detection methods, and providing an efficient and feasible new detection strategy for non-invasive early screening of bladder cancer.