TY - JOUR
T1 - LuNER
T2 - Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
AU - IGI Testing Consortium
AU - Stahl, Elizabeth C.
AU - Gopez, Allan R.
AU - Tsuchida, Connor A.
AU - Fan, Vinson B.
AU - Moehle, Erica A.
AU - Witkowsky, Lea B.
AU - Hamilton, Jennifer R.
AU - Lin-Shiao, Enrique
AU - McElroy, Matthew
AU - McDevitt, Shana L.
AU - Ciling, Alison
AU - Tsui, C. Kimberly
AU - Pestal, Kathleen
AU - Gildea, Holly K.
AU - Keller, Amanda
AU - Sylvain, Iman A.
AU - Williams, Clara
AU - Hirsh, Ariana
AU - Ehrenberg, Alexander J.
AU - Kantor, Rose
AU - Metzger, Matthew
AU - Nelson, Kara L.
AU - Urnov, Fyodor D.
AU - Ringeisen, Bradley R.
AU - Giannikopoulos, Petros
AU - Doudna, Jennifer A.
AU - Amen, Alexandra M.
AU - Barry, Kerrie W.
AU - Boyle, John M.
AU - Brook, Cara E.
AU - Choo, Seunga
AU - Cornmesser, L. T.
AU - Dilworth, David J.
AU - Fedrigo, Indro
AU - Friedline, Skyler E.
AU - Graham, Thomas G.W.
AU - Green, Ralph
AU - Hochstrasser, Megan L.
AU - Hockemeyer, Dirk
AU - Krishnappa, Netravathi
AU - Lari, Azra
AU - Li, Hanqin
AU - Lu, Tianlin
AU - Lyons, Elijah F.
AU - Mark, Kevin G.
AU - Martell, Lisa Argento
AU - Martins, Raquel O.A.
AU - Mitchell, Patrick S.
AU - Naca, Christine
AU - Nandakumar, Divya
N1 - Publisher Copyright:
© 2021 Stahl et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
PY - 2021/11
Y1 - 2021/11
N2 - Clinical and surveillance testing for the SARS-CoV-2 virus relies overwhelmingly on RTqPCR-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.
AB - Clinical and surveillance testing for the SARS-CoV-2 virus relies overwhelmingly on RTqPCR-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.
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U2 - 10.1371/journal.pone.0258263
DO - 10.1371/journal.pone.0258263
M3 - Article
C2 - 34758033
AN - SCOPUS:85120051765
SN - 1932-6203
VL - 16
JO - PloS one
JF - PloS one
IS - 11 November
M1 - e0258263
ER -