LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples.
oleh: Elizabeth C Stahl, Allan R Gopez, Connor A Tsuchida, Vinson B Fan, Erica A Moehle, Lea B Witkowsky, Jennifer R Hamilton, Enrique Lin-Shiao, Matthew McElroy, Shana L McDevitt, Alison Ciling, C Kimberly Tsui, Kathleen Pestal, Holly K Gildea, Amanda Keller, Iman A Sylvain, Clara Williams, Ariana Hirsh, Alexander J Ehrenberg, Rose Kantor, Matthew Metzger, Kara L Nelson, Fyodor D Urnov, Bradley R Ringeisen, Petros Giannikopoulos, Jennifer A Doudna, IGI Testing Consortium
| Format: | Article |
|---|---|
| Diterbitkan: | Public Library of Science (PLoS) 2021-01-01 |
Deskripsi
Clinical and surveillance testing for the SARS-CoV-2 virus relies overwhelmingly on RT-qPCR-based diagnostics, yet several popular assays require 2-3 separate reactions or rely on detection of a single viral target, which adds significant time, cost, and risk of false-negative results. Furthermore, multiplexed RT-qPCR tests that detect at least two SARS-CoV-2 genes in a single reaction are typically not affordable for large scale clinical surveillance or adaptable to multiple PCR machines and plate layouts. We developed a RT-qPCR assay using the Luna Probe Universal One-Step RT-qPCR master mix with publicly available primers and probes to detect SARS-CoV-2 N gene, E gene, and human RNase P (LuNER) to address these shortcomings and meet the testing demands of a university campus and the local community. This cost-effective test is compatible with BioRad or Applied Biosystems qPCR machines, in 96 and 384-well formats, with or without sample pooling, and has a detection sensitivity suitable for both clinical reporting and wastewater surveillance efforts.