Herein, a high-performance electrochemical sensor was constructed based on a metal-organic framework (Zr-UiO-66)/multi-carbon nanotubes/gold nanoparticles (Zr-UiO-66/MWCNTs/AuNPs) composite modified glassy carbon electrode for sensitive determination of tetracycline. The morphology, structure, and performance of Zr-UiO-66/MWCNTs/AuNPs were characterized by scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), energy dispersive spectroscopy (EDS), Fourier transform infrared spectroscopy (FT-IR), and electrochemical techniques. The Zr-UiO-66/MWCNTs/AuNPs nanohybrids exhibited excellent electrocatalytic activity towards the oxidation of tetracycline, mainly because of the synergistic effect of the MOFs, MWCNTs, and gold nanoparticles. The electrochemical kinetics and catalytical mechanism of tetracycline were demonstrated, proving that tetracycline's electrocatalytic oxidation reaction was an absorption-controlled two-step process involving the transfer of two protons and two electrons, respectively. Furthermore, a simple and facile method was used to achieve the regeneration of the absorbed saturated electrode. A low concentration of tetracycline was detected by amperometry with the linear ranges of 5 × 10-7 to 2.25 × 10-4 mol L-1 (R2 = 0.9941), the sensitivity was 45.4 mA L mol-1, and the limit of detection was as low as 1.67 × 10-7 mol L-1 (S/N = 3). In addition, the composite electrode demonstrated high selectivity (interference deviation of ± 5%), satisfactory reproductivity (RSD of 5.31%), and long-term stability (easy regeneration) and was successfully applied in the analysis of tetracycline antibiotics in actual samples. Thus, the proposed electrode provides a promising and prospective MOFs-based sensing platform for the detection of tetracyclines in the environment.