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In this work, the monodisperse polystyrene colloidal particles/Ag/zeolite imidazole framework (PS/Ag/ZIF-8) substrate was successfully prepared and served as SERS active substrate. The composition, structure and morphology of the PS/Ag/ZIF-8 substrates were studied by XRD, SEM, UV-Vis and XPS measurements. The main finding of this study was that the as-prepared PS/Ag/ZIF-8 substrate could exhibit outstanding SERS property when 4-mercaptobenzoic acid (4-MBA) was used as the SERS probes. The SERS mechanism was attributed to the combined