A Ni-MOF-based novel electrochemical sensor was synthesized with high surface area of 1381 m2/g, significant porosity of 1.14–9.6 nm and average particle size of 80 nm. The final products was developed for the detection of glucose with high sensitivity and accuracy. The amperometric response of the electrode toward glucose was achieved at a steady state in 3 s. The linear range, detection limit, and sensitivity of the developed electrode toward glucose was 1–1600 µM, 0.76 µM (S/N = 3) and 2859.95 µA mM−1 cm−2 (R2 = 0.9966), respectively. Compared with the non-enzymatic sensors, the developed electrode exhibited a lower detection limit and higher sensitivity, which are promising criteria for the development of a chemical sensor.