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Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method

Coronavirus Disease 2019 (COVID-19) caused by SARS-CoV-2 poses a significant threat to global public health. Early detection with reliable, fast, and simple assays is crucial to contain the spread of SARS-CoV-2. The real-time reverse transcription-polymerase chain reaction (RT-PCR) assay is currentl...

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Autores principales: Yang, Zhu, Liu, Nicole Y., Zhu, Zhiwei, Xiao, Minmin, Zhong, Shuzhi, Xue, Qiqi, Nie, Lina, Zhao, Jinhong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9558625/
https://www.ncbi.nlm.nih.gov/pubmed/36248705
http://dx.doi.org/10.7717/peerj.14121
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author Yang, Zhu
Liu, Nicole Y.
Zhu, Zhiwei
Xiao, Minmin
Zhong, Shuzhi
Xue, Qiqi
Nie, Lina
Zhao, Jinhong
author_facet Yang, Zhu
Liu, Nicole Y.
Zhu, Zhiwei
Xiao, Minmin
Zhong, Shuzhi
Xue, Qiqi
Nie, Lina
Zhao, Jinhong
author_sort Yang, Zhu
collection PubMed
description Coronavirus Disease 2019 (COVID-19) caused by SARS-CoV-2 poses a significant threat to global public health. Early detection with reliable, fast, and simple assays is crucial to contain the spread of SARS-CoV-2. The real-time reverse transcription-polymerase chain reaction (RT-PCR) assay is currently the gold standard for SARS-CoV-2 detection; however, the reverse transcription loop-mediated isothermal amplification method (RT-LAMP) assay may allow for faster, simpler and cheaper screening of SARS-CoV-2. In this study, the triple-target RT-LAMP assay was first established to simultaneously detect three different target regions (ORF1ab, N and E genes) of SARS-CoV-2. The results revealed that the developed triplex RT-LAMP assay was able to detect down to 11 copies of SARS-CoV-2 RNA per 25 µL reaction, with greater sensitivity than singleplex or duplex RT-LAMP assays. Moreover, two different indicators, hydroxy naphthol blue (HNB) and cresol red, were studied in the colorimetric RT-LAMP assay; our results suggest that both indicators are suitable for RT-LAMP reactions with an obvious color change. In conclusion, our developed triplex colorimetric RT-LAMP assay may be useful for the screening of COVID-19 cases in limited-resource areas.
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spelling pubmed-95586252022-10-14 Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method Yang, Zhu Liu, Nicole Y. Zhu, Zhiwei Xiao, Minmin Zhong, Shuzhi Xue, Qiqi Nie, Lina Zhao, Jinhong PeerJ Biochemistry Coronavirus Disease 2019 (COVID-19) caused by SARS-CoV-2 poses a significant threat to global public health. Early detection with reliable, fast, and simple assays is crucial to contain the spread of SARS-CoV-2. The real-time reverse transcription-polymerase chain reaction (RT-PCR) assay is currently the gold standard for SARS-CoV-2 detection; however, the reverse transcription loop-mediated isothermal amplification method (RT-LAMP) assay may allow for faster, simpler and cheaper screening of SARS-CoV-2. In this study, the triple-target RT-LAMP assay was first established to simultaneously detect three different target regions (ORF1ab, N and E genes) of SARS-CoV-2. The results revealed that the developed triplex RT-LAMP assay was able to detect down to 11 copies of SARS-CoV-2 RNA per 25 µL reaction, with greater sensitivity than singleplex or duplex RT-LAMP assays. Moreover, two different indicators, hydroxy naphthol blue (HNB) and cresol red, were studied in the colorimetric RT-LAMP assay; our results suggest that both indicators are suitable for RT-LAMP reactions with an obvious color change. In conclusion, our developed triplex colorimetric RT-LAMP assay may be useful for the screening of COVID-19 cases in limited-resource areas. PeerJ Inc. 2022-10-10 /pmc/articles/PMC9558625/ /pubmed/36248705 http://dx.doi.org/10.7717/peerj.14121 Text en ©2022 Yang et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Biochemistry
Yang, Zhu
Liu, Nicole Y.
Zhu, Zhiwei
Xiao, Minmin
Zhong, Shuzhi
Xue, Qiqi
Nie, Lina
Zhao, Jinhong
Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title_full Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title_fullStr Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title_full_unstemmed Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title_short Rapid and convenient detection of SARS-CoV-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
title_sort rapid and convenient detection of sars-cov-2 using a colorimetric triple-target reverse transcription loop-mediated isothermal amplification method
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9558625/
https://www.ncbi.nlm.nih.gov/pubmed/36248705
http://dx.doi.org/10.7717/peerj.14121
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