To sensitively monitor trace amounts of alternariol (AOH) in fruits, a dual-mode aptamer sensor utilizing the dual-function nanomaterial PoPD/Ru-Au was developed. This sensor provides both electrochemical (EC) and electrochemiluminescence (ECL) signals, which can greatly avoid the potential false positive of the traditional single signal, thus enhancing the accuracy and reliability of detection results. Polyo-phenylenediamine (PoPD), known for its favorable EC response, can also assist in enhancing the ECL behavior of Ru-Au. Furthermore, Ru-Au demonstrates excellent ECL performance and effectively activates K2S2O8 to amplify the EC response of PoPD. The complementary effect of the two can effectively amplify the final detection signal. Additionally, the PoPD/Ru-Au nanomaterial exhibits excellent electrical conductivity, further enhancing the EC and ECL response signals. The experimental results demonstrate that the EC detection range of AOH was 0.01-100 ng/mL, while the ECL detection range was 0.001-100 ng/mL, both exhibiting a satisfactory linear relationship. Therefore, the mutual verification of the detection results can be highly realized, and the purpose of avoiding wrong detection can be achieved.