The invention discloses a probe vector and a glucose probe and a preparation method and application thereof. The probe vector comprises a stainless steel needle and a nanogold layer wrapping the stainless steel needle; a three-dimensional continuous nano-channel structure is arranged on the surface of the nanogold layer. Compared with traditional porous gold prepared with pure gold line, gold foil and the like, the disposable nano-porous gold needle has the advantages of low cost, high mechanical strength, property of the porous gold, fitness for industrialized production and the like, and can be used as a microelectrode carrier of a biosensor that can be implanted in the human body at one step. Statistic indicate that conductivity and electrochemical active area of NiCo2O4 are improved, and glucose sensing property of the NiCo2O4 is thereby effectively improved.