An efficient surface-enhanced Raman scattering (SERS) substrate is developed based on silver nanoparticles decorated anodic aluminum oxide (Ag/AAO). The AAO templates were fabricated using a two-step anodization approach, and silver nanoparticles (AgNPs) were obtained by thermal decomposition of Ag nitrate in AAO. The structure of Ag/AAO hybrid substrate is characterized by scanning electron microscopy (SEM). The results show that the as-prepared SERS substrates consist of high-density AgNPs with sizes of tens of nanometers. The AgNPs are adsorbed on the surface of AAO template in the form of network structure which is called “hot spot”. The SERS enhancement ability of the nanostructure is verified using thiram as probing molecules. The limit of detection is as low as 1×10-9 mol/L. The results indicate that the as-prepared substrate possesses excellent SERS sensitivity, high stability and uniformity enhancement.