For developing highly selective and sensitive electrochemical sensors for chiral recognition, taking advantage of the synthetical properties of β-cyclodextrin (β-CD, strong host−guest recognition) and carbon nanotubes wrapped with reduced graphene oxide (CNTs@rGO, excellent electrochemical property and large surface area), as well as the differences in binding affinity between β-CD and guest molecules, a dual signal electrochemical sensing strategy was proposed herein for the first time in chiral recognition based on the competitive host−guest interaction between probe and chiral isomers with β-CD/CNTs@rGO. As a model system, rhodamine B (RhB) and phenylalanine enantiomers (Dand L-Phe) were introduced as probe and target enantiomers, respectively. Due to the host−guest interactions, RhB can enter into the β-CD cavity, showing remarkable oxidation peak current of RhB. In the presence of L-Phe, competitive interaction with the β-CD cavity occurs and RhB are replaced by L-Phe owing to the stronger binding affinity between L-Phe and β-CD, which results in the peak current of RhB decreasing and the peak current of L-Phe appears, and interestingly, the changes of both signals linearly correlate with the concentration of L-Phe. As for D-Phe, it cannot replace RhB owing to the weaker binding affinity between D-Phe and β-CD. Based on this, a dual-signal electrochemical sensor was developed successfully for recognizing Phe. This dual-signal sensing strategy can provide highly selective and sensitive recognition compared to single-signal sensor and has important potential applications in chiral recognition.C hirality has an important impact on chemical/biological research, since most active substances possess chirality. Generally, the performance of chiral enantiomers show great differences in terms of biochemical activity, toxicity, transport processes, and metabolic pathways. 1−3 Typically, only one isomer exhibits perfect activity and the other has no desirable value and even causes serious side-effects. Thus, chiral recognition is always a popular topic in chemical and biological research. 4−6 For resolving this problem, many approaches, such as capillary electrophoresis, 7 high-performance liquid chromatography, 8 circular dichroism spectroscopy, 9,10 colorimetry 11,12 and fluorescence, 13,14 were developed to recognize electroactive chiral molecules, which are very important in chiral recognition. However, the most of reported approaches need complex sample pretreatment and expensive chiral columns and are time-consuming as well. 15,16 Recently, electrochemical chiral recognition has received considerable attention, due to the many advantages they offer, such as low cost, fast response, inexpensive instrument, and facile miniaturization. 17−22 For instance, Kong et al. 23