LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples

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,...

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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8b3699807c8348d7aa2a3fafef2c5562
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spelling	oai:doaj.org-article:8b3699807c8348d7aa2a3fafef2c55622021-11-18T07:37:11ZLuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples1932-6203https://doaj.org/article/8b3699807c8348d7aa2a3fafef2c55622021-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8580221/?tool=EBIhttps://doaj.org/toc/1932-6203Clinical 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.Elizabeth C. StahlAllan R. GopezConnor A. TsuchidaVinson B. FanErica A. MoehleLea B. WitkowskyJennifer R. HamiltonEnrique Lin-ShiaoMatthew McElroyShana L. McDevittAlison CilingC. Kimberly TsuiKathleen PestalHolly K. GildeaAmanda KellerIman A. SylvainClara WilliamsAriana HirshAlexander J. EhrenbergRose KantorMatthew MetzgerKara L. NelsonFyodor D. UrnovBradley R. RingeisenPetros GiannikopoulosJennifer A. DoudnaIGI Testing ConsortiumPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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 LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
description	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.
format	article
author	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
author_facet	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
author_sort	Elizabeth C. Stahl
title	LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
title_short	LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
title_full	LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
title_fullStr	LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
title_full_unstemmed	LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples
title_sort	luner: multiplexed sars-cov-2 detection in clinical swab and wastewater samples
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/8b3699807c8348d7aa2a3fafef2c5562
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LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples

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