Objective To investigate the role of CA125, CA199 combined with exfoliative cytometer detection for the diagnosis of Ovarian Cancer. Methods 32 patients Ovarian Cancer with malignant seroperitoneum was detected. CA125 ,CA199 was detected with ELISA one-step method, and seroperitoneum exfoliative cytometer was detected for light microscope detection, the results were compared. Results Blood serum and seroperitoneum overall masculine response rate was 87.5%; CA199 detection overall masculine response rate was 28. 13%; seroperitoneum exfoliative cytometer detection overall masculine response rate was 53.13%;CA125, CA199 and exfoliative cytometer joint detection overall masculine response rate was 96.88%. There was significant difference about the overall response rate (P＜ 0. 05) between three groups, respectively. Conclusion Seroperitoneum tumor marker CA125, CA199 sensitivity is more height than simple seroperitoneum exfoliative cytometer detection, but Joint detection improve clinical diagnosis of ovarian cancer sensitivity.