Human epididymal protein 4(HE4), carbohydrate antigen 125 (CA125) and osteopontin(OPN) are three key biomarkers in detecting ovarian cancer.To explore the diagnosis value of combined detection of these three biomarkers for ovarian cancer, we developed a multiplexed assay on a plasmonic gold(pGOLD) platform for measuring HE4, CA125 and OPN in urine.The receiver operator characteristic(ROC) curve was drawn, and the diagnosis values of each biomarker alone or in combination for ovarian cancer were evaluated.In the anal. to distinguish ovarian cancer from other gynecol. cancers, ovarian cysts and healthy people, the sensitivities of HE4, CA125 and OPN were 72.55%, 52.82% and 68.63%, the specificity values were 95.06%, 87.65% and 90.12%, while the areas under the curve(AUC) were 0.85, 0.75 and 0.77, resp.The sensitivity and specificity for combination detection of the three markers were 90.20% and 80.25%.The detection methods of HE4, CA125 and OPN based on plasma fluorescence enhanced chip showed good analytic and diagnostic performance, and provided a non-invasive method for the diagnosis of ovarian cancer.