Surface-enhanced Raman scattering (SERS) spectroscopy has gained prominence in in vitro diagnostics due to its high sensitivity and multiplex detection capabilities. This review highlights recent advances in translational biomedical research using SERS techniques, with a focus on the rapid and accurate diagnosis of intractable diseases such as cancer and infectious diseases such as COVID-19. The review examines SERS-based assays for liquid biopsy biomarkers such as exosomes, circulating tumor cells, and circulating tumor DNAs that have emerged as promising tools in cancer diagnostics and are currently under active investigation in clinical research. Additionally, it explores SERS-based diagnostic techniques developed to overcome the limitations of existing COVID-19 diagnostic methods, including real-time polymerase chain reaction and lateral flow assay immunodiagnostics. Finally, the review addresses the challenges of translating SERS techniques from laboratory research to clinical practice.