Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2021 |
| Outros Autores: | , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | por |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/46841 |
Resumo: | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil. |
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Coelho, Bruna de OliveiraSanchuki, Heloisa Bruna SoligoZanette, Dalila LucíolaNardin, Jeanine MarieMorales, Hugo Manuel PazFornazari, BrunaAoki, Mateus NóbregaBlanes, Lucas2021-04-22T15:58:09Z2021-04-22T15:58:09Z2021COELHO, Bruna de Oliveira et al. Essential properties and pitfalls of colorimetric reverse transcription loop‑mediated isothermal amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis. Molecular Medicine, v.27, n. 30, p. 1–11, 2021.1528-3658https://www.arca.fiocruz.br/handle/icict/46841porBMCRT-LAMPAmplificação isotérmica mediada por loop de transcrição reversaSARS-CoV-2COVID-19Reverse-transcription loop-mediated isothermal amplifcationCoronavirus InfectionsReagent Kits, DiagnosticPoint-of-Care TestingInfecciones por CoronavirusJuego de Reactivos para DiagnósticoPruebas en el Punto de AtenciónInfections à coronavirusTrousses de réactifs pour diagnosticAnalyse sur le lieu d'interventionTranscrição ReversaInfecções por CoronavirusKit DiagnósticoTestes ImediatosEssential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.Hospital Erasto Gaertner. Curitiba, PR, Brasil.Hospital Erasto Gaertner. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil.SARS-CoV-2 Reverse Transcription Loop-mediated Isothermal Amplifcation (RT-LAMP) colorimetric detection is a sensitive and specifc point-of-care molecular biology technique used to detect the virus in only 30 min. In this manuscript we have described a few nuances of the technique still not properly described in the literature: the presence of three colors clusters; the correlation of the viral load with the color change; and the importance of using an internal control to avoid false-negative results. To achieve these fndings, we performed colorimetric RT-LAMP assays of 466 SARS-CoV-2 RT-qPCR validated clinical samples, with color quantifcation measured at 434 nm and 560 nm. First we determinate a sensitivity of 93.8% and specifcity of 90.4%. In addition to the pink (negative) and yellow (positive) produced colors, we report for the frst time the presence of an orange color cluster that may lead to wrong diagnosis. We also demonstrated using RT-qPCR and RT-LAMP that low viral loads are related to Ct values>30, resulting in orange colors. We also demonstrated that the diagnosis of COVID-19 by colorimetric RT-LAMP is efcient until the ffth symptoms day when the viral load is still relatively high. This study reports properties and indications for colorimetric RT-LAMP as point-of-care for SARS-CoV-2 diagnostic, reducing false results, interpretations and optimizing molecular diagnostics tests application.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/46841/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINAL10020_2021_Article_289ok.pdf10020_2021_Article_289ok.pdfapplication/pdf1227824https://www.arca.fiocruz.br/bitstream/icict/46841/2/10020_2021_Article_289ok.pdf90a7b80d78c2b42c405a066e875b9785MD52TEXT10020_2021_Article_289ok.pdf.txt10020_2021_Article_289ok.pdf.txtExtracted texttext/plain55627https://www.arca.fiocruz.br/bitstream/icict/46841/3/10020_2021_Article_289ok.pdf.txt1a1eb9397c440bf2fda887d2115d856cMD53icict/468412021-08-31 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| dc.title.pt_BR.fl_str_mv |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| title |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| spellingShingle |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis Coelho, Bruna de Oliveira RT-LAMP Amplificação isotérmica mediada por loop de transcrição reversa SARS-CoV-2 COVID-19 Reverse-transcription loop-mediated isothermal amplifcation Coronavirus Infections Reagent Kits, Diagnostic Point-of-Care Testing Infecciones por Coronavirus Juego de Reactivos para Diagnóstico Pruebas en el Punto de Atención Infections à coronavirus Trousses de réactifs pour diagnostic Analyse sur le lieu d'intervention Transcrição Reversa Infecções por Coronavirus Kit Diagnóstico Testes Imediatos |
| title_short |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| title_full |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| title_fullStr |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| title_full_unstemmed |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| title_sort |
Essential properties and pitfalls of colorimetric Reverse Transcription Loop‑mediated Isothermal Amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis |
| author |
Coelho, Bruna de Oliveira |
| author_facet |
Coelho, Bruna de Oliveira Sanchuki, Heloisa Bruna Soligo Zanette, Dalila Lucíola Nardin, Jeanine Marie Morales, Hugo Manuel Paz Fornazari, Bruna Aoki, Mateus Nóbrega Blanes, Lucas |
| author_role |
author |
| author2 |
Sanchuki, Heloisa Bruna Soligo Zanette, Dalila Lucíola Nardin, Jeanine Marie Morales, Hugo Manuel Paz Fornazari, Bruna Aoki, Mateus Nóbrega Blanes, Lucas |
| author2_role |
author author author author author author author |
| dc.contributor.author.fl_str_mv |
Coelho, Bruna de Oliveira Sanchuki, Heloisa Bruna Soligo Zanette, Dalila Lucíola Nardin, Jeanine Marie Morales, Hugo Manuel Paz Fornazari, Bruna Aoki, Mateus Nóbrega Blanes, Lucas |
| dc.subject.other.pt_BR.fl_str_mv |
RT-LAMP Amplificação isotérmica mediada por loop de transcrição reversa SARS-CoV-2 COVID-19 |
| topic |
RT-LAMP Amplificação isotérmica mediada por loop de transcrição reversa SARS-CoV-2 COVID-19 Reverse-transcription loop-mediated isothermal amplifcation Coronavirus Infections Reagent Kits, Diagnostic Point-of-Care Testing Infecciones por Coronavirus Juego de Reactivos para Diagnóstico Pruebas en el Punto de Atención Infections à coronavirus Trousses de réactifs pour diagnostic Analyse sur le lieu d'intervention Transcrição Reversa Infecções por Coronavirus Kit Diagnóstico Testes Imediatos |
| dc.subject.en.pt_BR.fl_str_mv |
Reverse-transcription loop-mediated isothermal amplifcation Coronavirus Infections Reagent Kits, Diagnostic Point-of-Care Testing |
| dc.subject.es.pt_BR.fl_str_mv |
Infecciones por Coronavirus Juego de Reactivos para Diagnóstico Pruebas en el Punto de Atención |
| dc.subject.fr.pt_BR.fl_str_mv |
Infections à coronavirus Trousses de réactifs pour diagnostic Analyse sur le lieu d'intervention |
| dc.subject.decs.pt_BR.fl_str_mv |
Transcrição Reversa Infecções por Coronavirus Kit Diagnóstico Testes Imediatos |
| description |
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Ciências e Tecnologias Aplicadas em Saúde. Curitiba, PR, Brasil. |
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2021 |
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2021-04-22T15:58:09Z |
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2021-04-22T15:58:09Z |
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2021 |
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COELHO, Bruna de Oliveira et al. Essential properties and pitfalls of colorimetric reverse transcription loop‑mediated isothermal amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis. Molecular Medicine, v.27, n. 30, p. 1–11, 2021. |
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https://www.arca.fiocruz.br/handle/icict/46841 |
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1528-3658 |
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COELHO, Bruna de Oliveira et al. Essential properties and pitfalls of colorimetric reverse transcription loop‑mediated isothermal amplification as a point‑of‑care test for SARS‑CoV‑2 diagnosis. Molecular Medicine, v.27, n. 30, p. 1–11, 2021. 1528-3658 |
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