The work aims to prepare carbon nanotube-nickel/boron-doped diamond composite electrode (CNTs-Ni/BDD) and use it in non-enzymatic glucose electrochemical detection. Hot filament chemical vapor deposition (HFCVD) was adopted to deposit BDD on silicon substrate and physical vapor deposition (PVD) was used to deposit Ni film on BDD. Then the Ni/BDD samples were put into a tube furnace for heat treatment at 900 ℃ for 30 min and 90 min respectively, to obtain CNTs-Ni/BDD electrodes with different microstructures. The surface morphology, composition and electrochemical performance of the films were characterized with scanning electron microscope (SEM), Raman spectroscope and electrochemical workstation, respectively. Finally, the CNTs grew from BDD directly with BDD as the substrate and the only carbon source at the high-temperature catalyst of Ni to modify BDD by Ni nano-particles and CNTs. In addition, if the heating time was increased from 30 min to 90 min, the length of CNTs increased significantly, the coverage degree on BDD improved and the Ni nano-particles disappeared. The joint modification of CNTs and Ni significantly improved the electrochemical detection performance of glucose. The 30 min-CNTs-Ni/BDD composite electrode showed a superior electrochemical performance and the sensitivity in the linear ranges of glucose concentration of 0.005~0.02 mmol/L, 0.02~1 mmol/L and 1.0~5.5 mmol/L was 475, 42, and 19 μA?(mmol/L)?1?cm?2, correspondingly. In addition, the detection limit was 0.42 μmol/L (S/N=3). The heat treatment can simply and effectively prepare a CNTs-Ni/BDD composite electrode which can effectively improve the electrochemical detection performance of glucose.