Recent advances in single-molecule detection, nanotechnology, and aptameric sensors hold exciting promise for many potential applications. By functionalizing the surface of a quantum dot (QD) with aptamers which can recognize cocaine, and taking advantage of single-molecule detection and fluorescence resonance energy transfer (FRET) between 605QD and Cy5 and Iowa Black RQ, we develop a single-QD-based aptameric sensor that is capable of sensing the presence of cocaine through both signal-off and signal-on modes. In comparison with the established aptameric sensors, this single-QD-based aptameric sensor has the significant advantages of simple sample preparation, high sensitivity, and extremely low sample consumption. With the advances in the development of varieties of aptamers for small molecules, nucleic acids, metal ions, and proteins, this single-QD-based aptameric sensor might find wide application in forensic analysis, environmental monitoring, and clinic diagnostics.