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/3053a4e5270e4f818e1a3cdef07b77fe
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spelling	oai:doaj.org-article:3053a4e5270e4f818e1a3cdef07b77fe2021-12-02T20:16:23ZLuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples.1932-620310.1371/journal.pone.0258263https://doaj.org/article/3053a4e5270e4f818e1a3cdef07b77fe2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0258263https://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, p e0258263 (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/3053a4e5270e4f818e1a3cdef07b77fe
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LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples.

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