Currently traumatic brain injury (TBI) is diagnosed with Glasgow Coma Scale (GCS), computed tomography (CT) scans or magnetic resonance imaging (MRI). Herein a paper-based optofluidic test strip has been developed to detect the TBI protein biomarker, neuron-specific enolase (NSE). A Surface-enhanced Raman scattering (SERS) substrate has been incorporated into a paper-based lateral flow strip. This paper-based device has achieved high sensitivity and low limit of detection in the blood plasma samples taken from the clinical TBI patients.