Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load

Abstract The use of RT-LAMP (reverse transcriptase—loop mediated isothermal amplification) has been considered as a promising point-of-care method to diagnose COVID-19. In this manuscript we show that the RT-LAMP reaction has a sensitivity of only 200 RNA virus copies, with a color change from pink...

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Autores principales:	Mateus Nóbrega Aoki, Bruna de Oliveira Coelho, Luiz Gustavo Bentim Góes, Paola Minoprio, Edison Luiz Durigon, Luis Gustavo Morello, Fabricio Klerynton Marchini, Irina Natassja Riediger, Maria do Carmo Debur, Helder I. Nakaya, Lucas Blanes
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b0e312b342174d3399d2188b1de467c5
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spelling	oai:doaj.org-article:b0e312b342174d3399d2188b1de467c52021-12-02T13:40:51ZColorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load10.1038/s41598-021-88506-y2045-2322https://doaj.org/article/b0e312b342174d3399d2188b1de467c52021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-88506-yhttps://doaj.org/toc/2045-2322Abstract The use of RT-LAMP (reverse transcriptase—loop mediated isothermal amplification) has been considered as a promising point-of-care method to diagnose COVID-19. In this manuscript we show that the RT-LAMP reaction has a sensitivity of only 200 RNA virus copies, with a color change from pink to yellow occurring in 100% of the 62 clinical samples tested positive by RT-qPCR. We also demonstrated that this reaction is 100% specific for SARS-CoV-2 after testing 57 clinical samples infected with dozens of different respiratory viruses and 74 individuals without any viral infection. Although the majority of manuscripts recently published using this technique describe only the presence of two-color states (pink = negative and yellow = positive), we verified by naked-eye and absorbance measurements that there is an evident third color cluster (orange), in general related to positive samples with low viral loads, but which cannot be defined as positive or negative by the naked eye. Orange colors should be repeated or tested by RT-qPCR to avoid a false diagnostic. RT-LAMP is therefore very reliable for samples with a RT-qPCR Ct < 30 being as sensitive and specific as a RT-qPCR test. All reactions were performed in 30 min at 65 °C. The use of reaction time longer than 30 min is also not recommended since nonspecific amplifications may cause false positives.Mateus Nóbrega AokiBruna de Oliveira CoelhoLuiz Gustavo Bentim GóesPaola MinoprioEdison Luiz DurigonLuis Gustavo MorelloFabricio Klerynton MarchiniIrina Natassja RiedigerMaria do Carmo DeburHelder I. NakayaLucas BlanesNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Mateus Nóbrega Aoki Bruna de Oliveira Coelho Luiz Gustavo Bentim Góes Paola Minoprio Edison Luiz Durigon Luis Gustavo Morello Fabricio Klerynton Marchini Irina Natassja Riediger Maria do Carmo Debur Helder I. Nakaya Lucas Blanes Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
description	Abstract The use of RT-LAMP (reverse transcriptase—loop mediated isothermal amplification) has been considered as a promising point-of-care method to diagnose COVID-19. In this manuscript we show that the RT-LAMP reaction has a sensitivity of only 200 RNA virus copies, with a color change from pink to yellow occurring in 100% of the 62 clinical samples tested positive by RT-qPCR. We also demonstrated that this reaction is 100% specific for SARS-CoV-2 after testing 57 clinical samples infected with dozens of different respiratory viruses and 74 individuals without any viral infection. Although the majority of manuscripts recently published using this technique describe only the presence of two-color states (pink = negative and yellow = positive), we verified by naked-eye and absorbance measurements that there is an evident third color cluster (orange), in general related to positive samples with low viral loads, but which cannot be defined as positive or negative by the naked eye. Orange colors should be repeated or tested by RT-qPCR to avoid a false diagnostic. RT-LAMP is therefore very reliable for samples with a RT-qPCR Ct < 30 being as sensitive and specific as a RT-qPCR test. All reactions were performed in 30 min at 65 °C. The use of reaction time longer than 30 min is also not recommended since nonspecific amplifications may cause false positives.
format	article
author	Mateus Nóbrega Aoki Bruna de Oliveira Coelho Luiz Gustavo Bentim Góes Paola Minoprio Edison Luiz Durigon Luis Gustavo Morello Fabricio Klerynton Marchini Irina Natassja Riediger Maria do Carmo Debur Helder I. Nakaya Lucas Blanes
author_facet	Mateus Nóbrega Aoki Bruna de Oliveira Coelho Luiz Gustavo Bentim Góes Paola Minoprio Edison Luiz Durigon Luis Gustavo Morello Fabricio Klerynton Marchini Irina Natassja Riediger Maria do Carmo Debur Helder I. Nakaya Lucas Blanes
author_sort	Mateus Nóbrega Aoki
title	Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
title_short	Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
title_full	Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
title_fullStr	Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
title_full_unstemmed	Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load
title_sort	colorimetric rt-lamp sars-cov-2 diagnostic sensitivity relies on color interpretation and viral load
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b0e312b342174d3399d2188b1de467c5
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Colorimetric RT-LAMP SARS-CoV-2 diagnostic sensitivity relies on color interpretation and viral load

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