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IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2

Commonly used RT-qPCR-based SARS-CoV-2 diagnostics require 2–3 separate reactions or rely on detection of a single viral target, adding time and cost or risk of false-negative results. Currently, no test combines detection of widely used SARS-CoV-2 E- and N-gene targets and a sample control in a sin...

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Autores principales: Stahl, Elizabeth C., Tsuchida, Connor A., Hamilton, Jennifer R., Lin-Shiao, Enrique, McDevitt, Shana L., Moehle, Erica A., Witkowsky, Lea B., Tsui, C. Kimberly, Pestal, Kathleen, Gildea, Holly K., McElroy, Matthew, Keller, Amanda, Sylvain, Iman, Williams, Clara, Hirsh, Ariana, Ciling, Alison, Ehrenberg, Alexander J., Urnov, Fyodor D., Ringeisen, Bradley R., Giannikopoulos, Petros, Doudna, Jennifer A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7743092/
https://www.ncbi.nlm.nih.gov/pubmed/33330883
http://dx.doi.org/10.1101/2020.12.10.20247338
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author Stahl, Elizabeth C.
Tsuchida, Connor A.
Hamilton, Jennifer R.
Lin-Shiao, Enrique
McDevitt, Shana L.
Moehle, Erica A.
Witkowsky, Lea B.
Tsui, C. Kimberly
Pestal, Kathleen
Gildea, Holly K.
McElroy, Matthew
Keller, Amanda
Sylvain, Iman
Williams, Clara
Hirsh, Ariana
Ciling, Alison
Ehrenberg, Alexander J.
Urnov, Fyodor D.
Ringeisen, Bradley R.
Giannikopoulos, Petros
Doudna, Jennifer A.
author_facet Stahl, Elizabeth C.
Tsuchida, Connor A.
Hamilton, Jennifer R.
Lin-Shiao, Enrique
McDevitt, Shana L.
Moehle, Erica A.
Witkowsky, Lea B.
Tsui, C. Kimberly
Pestal, Kathleen
Gildea, Holly K.
McElroy, Matthew
Keller, Amanda
Sylvain, Iman
Williams, Clara
Hirsh, Ariana
Ciling, Alison
Ehrenberg, Alexander J.
Urnov, Fyodor D.
Ringeisen, Bradley R.
Giannikopoulos, Petros
Doudna, Jennifer A.
author_sort Stahl, Elizabeth C.
collection PubMed
description Commonly used RT-qPCR-based SARS-CoV-2 diagnostics require 2–3 separate reactions or rely on detection of a single viral target, adding time and cost or risk of false-negative results. Currently, no test combines detection of widely used SARS-CoV-2 E- and N-gene targets and a sample control in a single, multiplexed reaction. We developed the IGI-LuNER 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 (NER). This combined, cost-effective test can be performed in 384-well plates with detection sensitivity suitable for clinical reporting, and will aid in future sample pooling efforts, thus improving throughput of SARS-CoV-2 detection.
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spelling pubmed-77430922020-12-17 IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2 Stahl, Elizabeth C. Tsuchida, Connor A. Hamilton, Jennifer R. Lin-Shiao, Enrique McDevitt, Shana L. Moehle, Erica A. Witkowsky, Lea B. Tsui, C. Kimberly Pestal, Kathleen Gildea, Holly K. McElroy, Matthew Keller, Amanda Sylvain, Iman Williams, Clara Hirsh, Ariana Ciling, Alison Ehrenberg, Alexander J. Urnov, Fyodor D. Ringeisen, Bradley R. Giannikopoulos, Petros Doudna, Jennifer A. medRxiv Article Commonly used RT-qPCR-based SARS-CoV-2 diagnostics require 2–3 separate reactions or rely on detection of a single viral target, adding time and cost or risk of false-negative results. Currently, no test combines detection of widely used SARS-CoV-2 E- and N-gene targets and a sample control in a single, multiplexed reaction. We developed the IGI-LuNER 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 (NER). This combined, cost-effective test can be performed in 384-well plates with detection sensitivity suitable for clinical reporting, and will aid in future sample pooling efforts, thus improving throughput of SARS-CoV-2 detection. Cold Spring Harbor Laboratory 2020-12-11 /pmc/articles/PMC7743092/ /pubmed/33330883 http://dx.doi.org/10.1101/2020.12.10.20247338 Text en https://creativecommons.org/licenses/by-nd/4.0/This work is licensed under a Creative Commons Attribution-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, and only so long as attribution is given to the creator. The license allows for commercial use.
spellingShingle Article
Stahl, Elizabeth C.
Tsuchida, Connor A.
Hamilton, Jennifer R.
Lin-Shiao, Enrique
McDevitt, Shana L.
Moehle, Erica A.
Witkowsky, Lea B.
Tsui, C. Kimberly
Pestal, Kathleen
Gildea, Holly K.
McElroy, Matthew
Keller, Amanda
Sylvain, Iman
Williams, Clara
Hirsh, Ariana
Ciling, Alison
Ehrenberg, Alexander J.
Urnov, Fyodor D.
Ringeisen, Bradley R.
Giannikopoulos, Petros
Doudna, Jennifer A.
IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title_full IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title_fullStr IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title_full_unstemmed IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title_short IGI-LuNER: single-well multiplexed RT-qPCR test for SARS-CoV-2
title_sort igi-luner: single-well multiplexed rt-qpcr test for sars-cov-2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7743092/
https://www.ncbi.nlm.nih.gov/pubmed/33330883
http://dx.doi.org/10.1101/2020.12.10.20247338
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