In the present paper, quantum chemistry calculations method based on the density functional theory (DFT) and surface-enhanced Raman scattering (SERS) spectroscopy technique were used to investigate the adsorption behavior and enhancement effect of thiabendazole on the nanometer silver colloid surface systematically from theoretical and experimental perspective. By sodium citrate’s reduction reaction, nanometer silver colloid with has high surface-enhanced Raman scattering activity was prepared. And then the authors studied the surface-enhanced Raman scattering spectroscopy of the thiabendazole in aqueous solution. The authors carried on the detailed quantum chemistry calculations for the interaction between thiabendazole and nanometer silver colloid, using the TBZ-Ag4 model to get the adsorption properties of thiabendazole molecule on nanometer silver colloid. Combining FT-Raman spectrum with the theoretical calculation results by the B3LYP/6-311G(d) theoretical level, and the visualization of GaussianView5.0 software, the FT-Raman vibration spectrum and the surface-enhanced Raman scattering spectroscopy of thiabendazole molecule were assigned systematically. All the theoretical and experimental results show that all atoms of thiabendazole are in one plane and the point group of thiabendazole is Cs; Thiabendazole has high surface-enhanced Raman scattering activity on nanometer silver colloid surface; the thiabendazole is absorbed on silver colloid particles by S atom, and the long axis of thiabendazole molecule is perpendicular to the nanometer silver colloid surface; the trace concentration of thiabendazole can be detected rapidly and effectively with the surface-enhanced Raman scattering spectroscopy technique. This work provides a theoretical and experimental basis for the study of thiabendazole’s characteristics and its rapid detection.