Rapid and sensitive diagnostics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of utmost importance to control the widespread coronavirus disease 2019 (COVID-19) upsurge. This study demonstrated a novel one-pot surface-enhanced Raman scattering (SERS) based immunoassay to detect SARS-CoV-2, without any washing process using a portable Raman spectrometer. The SERS-immune assay was designed using a regular digital versatile disk (DVD) substrate integrated with Raman reporter labeled silver nanoparticles for double clamping effects. The disks were molded to form nanopillar arrays and coated with silver film to enhance the sensitivity of immunoassay. The SERS platform demonstrated a limit of detection (LoD) up to 50 pg mL−1 for SARS-CoV-2 spike protein and virus-like-particle (VLP) protein in phosphate buffer saline within a turnaround time of 20 mins. Moreover, VLP protein spiked in untreated saliva achieved an LoD of 400 pg mL−1, providing a cycle threshold (Ct) value range of 30–32, closer to reverse transcription-polymerase chain reaction (RT-PCR) results (35−40) and higher than the commercial rapid antigen tests, ranging from 25 to 28. Therefore, the developed one-pot SERS based biosensor exhibited highly sensitive and rapid detection of SARS-CoV-2, which could be a potential point-of-care platform for early and cost-effective diagnosis of the COVID-19 virus. Display omitted •Dual-clamped One-Pot SERS biosensor for rapid and highly sensitive detection of SARS-CoV-2.•One-Pot approach without washing process demonstrated a turnaround time of 20 mins by Portable Raman spectrometer.•Regular DVD discs as the SERS substrate improves the cost and reproducibility of SERS immunoassay.•LoDs of SARS-CoV-2 VLP protein obtained were 50 pg mL−1 in PBS and 400 pg mL−1 in untreated saliva respectively.•LoD enhancement of more than 100 times achieved compared to commercial antigen tests.