A novel dual-mode cholesterol biosensor was developed based on gold nanorods-carbon quantum dots-Cholesterol oxidase (AuNRs-CQDs-COD) complex synthesized by the cross-linking method. The specific reaction to/against cholesterol and cholesterol oxidase can produce hydrogen peroxide. Due to the etching effect between the hydrogen peroxide and AuNRs, the absorption peaks of AuNRs would drift so that the fluorescence quenching occurred because of the electronic charge transfer between the hydrogen peroxide and CQDs. The spectrum properties, structure and micromorphology of AuNRs-CQDs-COD complex were characterized by transmission electron microscope (TEM), Fourier transform infrared (FTIR), Fluorescence and Ultraviolet–Visible absorption spectroscopy, respectively. There was a good linear correlation between the fluorescence response and absorption peak shift of AuNRs-CQDs-COD probe in cholesterol solution within the concentration in the range 0.1–1 mM, and the limit of detection (LOD) is 0.028 μM. This dual-mode cholesterol biosensor based on AuNRs-CQDs-COD complex has good anti-interference and selectivity.