Early detection of ovarian cancer remains a significant challenge due to the lack of symptoms in its early stages and the overlap in protein expression patterns between malignant and benign conditions. In this study, we introduce a dual gatekeepers-driven signal amplification strategy for the highly sensitive detection and precise modulation of ovarian cancer-derived exosome subtypes. By employing CA125 and carcinoembryonic antigen (CEA) aptamers as dual gatekeepers, this strategy selectively activates functional regions on exosome membranes, triggering the opening of hairpin DNA structures (HP) upon recognition of specific protein patterns. The opened HP then initiate an exonuclease III-powered DNA walking system and nucleic acid-stabilized Ag