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Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2

A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplificat...

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Autores principales: Jang, Woong Sik, Lim, Da Hye, Yoon, Jung, Kim, Ahran, Lim, Minsup, Nam, Jeonghun, Yanagihara, Richard, Ryu, Sook-Won, Jung, Bo Kyeung, Ryoo, Nam-Hee, Lim, Chae Seung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7928493/
https://www.ncbi.nlm.nih.gov/pubmed/33657176
http://dx.doi.org/10.1371/journal.pone.0248042
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author Jang, Woong Sik
Lim, Da Hye
Yoon, Jung
Kim, Ahran
Lim, Minsup
Nam, Jeonghun
Yanagihara, Richard
Ryu, Sook-Won
Jung, Bo Kyeung
Ryoo, Nam-Hee
Lim, Chae Seung
author_facet Jang, Woong Sik
Lim, Da Hye
Yoon, Jung
Kim, Ahran
Lim, Minsup
Nam, Jeonghun
Yanagihara, Richard
Ryu, Sook-Won
Jung, Bo Kyeung
Ryoo, Nam-Hee
Lim, Chae Seung
author_sort Jang, Woong Sik
collection PubMed
description A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer sets to detect the RNA-dependent RNA polymerase (RdRP), Envelope (E) and Nucleocapsid protein (N) genes of SARS CoV-2. For one tube reaction, the detection limits for five combination SARS CoV-2 LAMP primer sets (RdRP/E, RdRP/N, E/N, RdRP/E/N and RdRP/N/Internal control (actin beta)) were evaluated with a clinical nasopharyngeal swab sample. Among the five combination, the RdRP/E and RdRP/N/IC multiplex LAMP assays showed low detection limits. The sensitivity and specificity of the RT-LAMP assay were evaluated and compared to that of the widely used Allplex™ 2019-nCoV Assay (Seegene, Inc., Seoul, South Korea) and PowerChek™ 2019-nCoV Real-time PCR kit (Kogenebiotech, Seoul, South Korea) for 130 clinical samples from 91 SARS CoV-2 patients and 162 NP specimens from individuals with (72) and without (90) viral respiratory infections. The multiplex RdRP (FAM)/N (CY5)/IC (Hex) RT-LAMP assay showed comparable sensitivities (RdRP: 93.85%, N: 94.62% and RdRP/N: 96.92%) to that of the Allplex™ 2019-nCoV Assay (100%) and superior to those of PowerChek™ 2019-nCoV Real-time PCR kit (RdRP: 92.31%, E: 93.85% and RdRP/E: 95.38%).
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spelling pubmed-79284932021-03-10 Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2 Jang, Woong Sik Lim, Da Hye Yoon, Jung Kim, Ahran Lim, Minsup Nam, Jeonghun Yanagihara, Richard Ryu, Sook-Won Jung, Bo Kyeung Ryoo, Nam-Hee Lim, Chae Seung PLoS One Research Article A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer sets to detect the RNA-dependent RNA polymerase (RdRP), Envelope (E) and Nucleocapsid protein (N) genes of SARS CoV-2. For one tube reaction, the detection limits for five combination SARS CoV-2 LAMP primer sets (RdRP/E, RdRP/N, E/N, RdRP/E/N and RdRP/N/Internal control (actin beta)) were evaluated with a clinical nasopharyngeal swab sample. Among the five combination, the RdRP/E and RdRP/N/IC multiplex LAMP assays showed low detection limits. The sensitivity and specificity of the RT-LAMP assay were evaluated and compared to that of the widely used Allplex™ 2019-nCoV Assay (Seegene, Inc., Seoul, South Korea) and PowerChek™ 2019-nCoV Real-time PCR kit (Kogenebiotech, Seoul, South Korea) for 130 clinical samples from 91 SARS CoV-2 patients and 162 NP specimens from individuals with (72) and without (90) viral respiratory infections. The multiplex RdRP (FAM)/N (CY5)/IC (Hex) RT-LAMP assay showed comparable sensitivities (RdRP: 93.85%, N: 94.62% and RdRP/N: 96.92%) to that of the Allplex™ 2019-nCoV Assay (100%) and superior to those of PowerChek™ 2019-nCoV Real-time PCR kit (RdRP: 92.31%, E: 93.85% and RdRP/E: 95.38%). Public Library of Science 2021-03-03 /pmc/articles/PMC7928493/ /pubmed/33657176 http://dx.doi.org/10.1371/journal.pone.0248042 Text en © 2021 Jang et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Jang, Woong Sik
Lim, Da Hye
Yoon, Jung
Kim, Ahran
Lim, Minsup
Nam, Jeonghun
Yanagihara, Richard
Ryu, Sook-Won
Jung, Bo Kyeung
Ryoo, Nam-Hee
Lim, Chae Seung
Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title_full Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title_fullStr Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title_full_unstemmed Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title_short Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2
title_sort development of a multiplex loop-mediated isothermal amplification (lamp) assay for on-site diagnosis of sars cov-2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7928493/
https://www.ncbi.nlm.nih.gov/pubmed/33657176
http://dx.doi.org/10.1371/journal.pone.0248042
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