We report on a synthetic approach to produce self-supported flexible surface-enhanced Raman scattering (SERS) active membranes consisting of polyamide (PA) nanofibers grafted with vertical Ag-nanosheets, via a combinatorial process of electrospinning PA-nanofiber membranes, assembling Au-nanoparticles on the PA-nanofibers as seeds for subsequent growth of Ag-nanosheets, and electrodepositing Ag-nanosheets on the electrospun PA-nanofibers. As a high density of Ag-nanosheets are vertically grown around each PA-nanofiber in the three-dimensional (3D) networked PA-nanofiber membranes, homogeneous nano-scaled gaps between the neighboring Ag-nanosheets are formed, leading to a high density of 3D SERS "hot spots" within the Ag-nanosheet-grafted PA-nanofiber membranes. The Ag-nanosheet-grafted PA-nanofiber membranes demonstrate high SERS activity with excellent Raman signal reproducibility for rhodamine 6G over the whole membrane. For a SERS-based trial analysis of polychlorinated biphenyls (PCBs, a kind of global environmental hazard), the 3D SERS substrate membranes are modified with mono-6-β-cychlodextrin to effectively capture PCB molecules. As a result, not only a low concentration down to 10(-6) M is reached, but also two congeners of PCBs in their mixed solution are identified, showing promising potential in SERS-based rapid detection of trace organic pollutants such as PCBs in the environment.