A high-efficiency surface-enhanced Raman scattering (SERS) detection method with ultra-high sensitivity has been widely applied in drug component detection to optimize the product quality verification standards. Herein, a controllable strategy of sputtering Ag nanoislands on carbon fiber (C-fiber) via magnetron sputtering technology was proposed to fabricate a versatile Ag-C-fiber SERS active substrate. A wide range of multi-level electromagnetic enhancement “hot spots” distributed on Ag-C-fiber nanostructures can efficiently amplify Raman signals and the experimental enhancement factor (EEF) value was 3.871 × 10⁶. Furthermore, substantial “hot spots” of large-scale distribution guaranteed the superior reproducibility of Raman signal with relative standard deviation (RSD) values less than 12.97%. Limit of detection (LOD) results indicated that when crystal violet (CV) is employed as probe molecule, the LOD was located at 1 × 10⁻¹³ M. By virtue of ultra-sensitivity and good flexibility of the Ag-C-fiber nanotemplate, Raman signals of two kinds of antiepileptic drugs called levetiracetam and sodium valproate were successfully obtained using an SERS-based spectral method. The Ag-C-fiber SERS detection platform demonstrated a good linear response (R² = 0.97486) in sensing sodium valproate concentrations in the range of 1 × 10³ ng/μL⁻¹–1 ng/μL. We believe that this reliable strategy has potential application for trace detection and rapid screening of antiepileptic drugs in the clinic.