SARS-CoV-2 has created a global disease burden infecting >100 million humans in just over a year. In this study, we adopted a SISCAPA-based enrichment approach using anti-peptide antibodies generated against peptides from the nucleocapsid protein of SARS-CoV-2. We developed a targeted workflow in which nasopharyngeal samples were digested followed by enrichment of viral peptides using the anti-peptide antibodies and targeted parallel reaction monitoring (PRM) analysis using a high-resolution mass spectrometer. This workflow was applied to 41 RT-PCR-confirmed clinical nasopharyngeal swab samples and 30 negative samples. The SISCAPA-based platform described in the current study can serve as one of the alternative methods for SARS-CoV-2 viral detection.