A flexible surface-enhanced Raman scattering (SERS) substrate was prepared by vacuum evaporation of silver on the surface of woven nylon fabrics. SERS properties of the Ag-coated nylon fabrics varied as the thickness of silver coatings changed, relative to the morphologies and distribution of silver nanoparticles (NPs) on fiber. The SERS enhancement performance of Ag-coated nylon fabrics was evaluated by collecting Raman signals of different concentrations of p-aminothiophenol (PATP). The results suggested that the nylon fabrics coated with 10 nm thickness Ag NPs coatings possessed high SERS activity and its detection concentration for PATP is as low as 10−9 M. Furthermore, sensitive SERS signals with excellent reproducibility (Relative standard deviation = 8.25%) and stability (30 days) have been demonstrated. In addition, the SERS nylon fabrics have been applied to rapidly detect thiram pesticides on cucumber, which indicated a great potential for trace analysis.