Rapid detection of SARS-CoV-2 variants by molecular-clamping technology-based RT-qPCR
We have developed a multiplex reverse-transcription quantitative real-time PCR (RT-qPCR) testing platform for the rapid detection of SARS-CoV-2 variants using the xenonucleic acid (XNA)-based molecular-clamping technology. The XNA-based RT-qPCR assay can achieve high sensitivity with a limit of detection of about 100 copies/mL for variant detection which is much better than the next-generation sequencing (NGS) assay. Its turnaround time is about 4 hours with lower cost and a lot of Clinical Laboratory Improvement Amendments (CLIA) labs own the instrument and meet skillset requirements. This assay provides a rapid, reliable, and cost-effective testing platform for rapid detection and monitoring of known and emerging SARS-CoV-2 variants. This testing platform can be adopted by laboratories that perform routine SARS-CoV-2 PCR testing, providing a rapid and cost-effective method in lieu of NGS-based assays, for detecting, differentiating, and monitoring SARS-CoV-2 variants. This assay is easily scalable to any new variant(s) should it emerge.
Authors
- Daniel B. Kissinger
- Joseph A. Pathakamuri
- Larry Pastor
- Daniel Kuebler (ORCID: https://orcid.org/0000-0001-8641-8149)
- Daniel Cataldi
- Isaac Archibeque
- Jonathan Z. Li (ORCID: https://orcid.org/0000-0001-9914-9662)
- Shuo Shen (ORCID: https://orcid.org/0000-0002-3517-8971)
- Michael Rohall
- Chuanyi Mark Lu (ORCID: https://orcid.org/0000-0002-5906-2543)
- Jocelyn Neves
- Maidar Jamba
- Madison Krohn
- Andrew Y. Fu (ORCID: https://orcid.org/0000-0002-1507-0787)
- Michael Y. Sha (ORCID: https://orcid.org/0000-0002-4552-6079)
- Qing Sun (ORCID: https://orcid.org/0000-0001-5158-1106)
- Aiguo Zhang (ORCID: https://orcid.org/0000-0002-6596-1895)
- Zhen Cui (ORCID: https://orcid.org/0000-0001-8723-1175)
Institutions
- Franciscan University of Steubenville (US)
- University of California, San Francisco (US)
- San Francisco VA Health Care System (US)
Publication Details
- Journal
- Microbiology Spectrum
- Published
- 2024-10-16
- DOI
- https://doi.org/10.1128/spectrum.04248-23
- Citations
- 2
- Primary Topic
- SARS-CoV-2 detection and testing
- Type
- article
- Field-Weighted Citation Impact
- 0.78
Funders
- University of California, San Francisco