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The β-nickel sulfide (β-NiS) nanobelts were fabricated by electrodepositing a nickel nanosheet film on Indium tin oxide (ITO)-coated glass substrates and sulfuring the nickel film on ITO-coated glass substrates. The sulfurization method can be used to form nanobelts without a template. A small glass tube was used to anneal the sulfur sheet with a nickel nanosheet film. After applying vacuum to the tube, the specimen was annealed at 500 °C. By adjusting the weight of the sulfur sheet in a small glass tube, a nanobelt structure can be formed on the film for 4 h. The β-NiS nanobelt film had a sulfide and nickel molar ratio that was nearly 0.7 (S/Ni). After five years of a long-term storage test, the β-NiS nanobelt films were able to measure the glucose in a solution with the value of sensitivity of 8.67 µA cm−2 µM−1. The β-NiS nanobelt film also detected glucose with a limit of low detection (LOD) of around 0.173 µM. The estimation of reproducibility was over 98%. Therefore, the β-NiS nanobelt film has a significant ability to detect low concentrations of glucose in a solution.