The key to developing sensors for chiral drug determination is to exclude interference from enantiomers. In this study, metal-organic frameworks (MOFs) and molecularly imprinted polymer (MIP) were introduced to prepare a chiral sensor for levofloxacin detection. The MIP was electropolymerised on the surface of the Cu/Fe-benzene-1,3,5-tricarboxylate MOF (Cu/Fe-BTC)-modified Au electrode using levofloxacin as a template molecule. After eluting the levofloxacin, a chiral sensor with recognition sites for levofloxacin was obtained. With this site as a switch, a novel method for detecting levofloxacin was established. Because of the enhanced recognition effect, the sensor can effectively exclude the enantiomeric interference of d-ofloxacin. Moreover, Cu/Fe-BTC can effectively amplify the current response signal and improve the sensitivity of thesensor. The linear range of the sensor was 5 to 4000 × 10-11mol L-1, and the detection limit was 2.07 × 10-11mol L-1. Whenapplied to detecting levofloxacin in actual samples, the sensor showed a 92.7-109.8% recovery.