Au nanorods were prepared by a seeding growth approach and used in fabricating the nanorod-enhancing glucose biosensor. The high affinity of chitosan for Au nanorods associated with its amino groups resulted in the formation of a layer of Au nanorods on the surface of Au electrode. It served as an intermediator to retain high efficient and stable immobilization of the enzyme. The performance of biosensors was investigated by cyclic voltammetry (CV), in the presence of artificial redox mediator, ferrocenecarboxaldehyde. The biosensor had a fast response to glucose, and the response time was less than 10 s. The results indicated that the gold nanorods could enhance the current response to glucose. The detection limits of glucose can reach 10 mM, and the Michaelis-Menten constant Kappm 13.62 mM.