In this study, the electrochemical cyclic voltammetry method was developed for the determination of activity of glucose oxidase. The ferrocene methanol was introducing as the redox probes on cyclic voltammetry, using glassy carbon electrode as working electrode, saturated calomel electrode as reference electrode, platinum wire electrode as the counter electrode and the peak current value of different enzyme activity was required . There was a linear relationship between the peak current in enzyme concentration of 1.282～2.377 U/mL with the correlation coefficient as 0.998, the detection limit as 1.282 U/mL. The enzyme activity of spores from dormancy to germination was detected by this new method, as the enzyme activity of spores decreased after increasing first, reaching the maximum (2.130 U/mL). The traditional methods were tested at the same time. Compared with the traditional method, the proposed electrochemical method has the characteristics of accurate, fast and convenient.