The present disclosure relates to a method for manufacturing a working electrode of a glucose monitoring probe, comprising: preparing a substrate; forming a base layer on the substrate; preparing a sensing layer reagent comprising a redoxpolymer, a glucose enzyme, a carbon nanotube, and a cross-linking agent, adding an amino modification to the carbon nanotube to enable the redoxpolymer and the carbon nanotube to form a covalent bond, and combining the carbon nanotube with the glucose enzyme, wherein the working voltage required by the working electrode is reduced under the catalytic action of the carbon nanotube, and the glucose enzyme is glucose oxidase; coating the sensing layer reagent on the base layer to form a sensing layer; forming a semi-permeable membrane on the sensing layer to control the passage rate of glucose molecules; and forming a biocompatible membrane on the semi-permeable membrane. According to the present disclosure, a method for manufacturing a working electrode of a glucose monitoring probe capable of reducing the working voltage of the working electrode and improving the reaction sensitivity to glucose can be provided.