Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2021 |
| Outros Autores: | , , , , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/51705 |
Resumo: | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil. |
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Alves, Pedro AugustoOliveira, Ellen Gonçalves deLuiz, Ana Paula Moreira FrancoAlmeida, Letícia TrindadeGonçalves, Amanda BonotoBorges, Iara ApolinárioRocha, Flávia de SRocha, Raissa PradoBezerra, Matheus FilgueiraMiranda, PâmellaCapanema, Flávio DinizMartins, Henrique ResendeWeber, GeraldTeixeira, Santuza Maria RibeiroWallau, Gabriel LuzMonte Neto, Rubens Lima do2022-03-16T19:30:39Z2022-03-16T19:30:39Z2021ALVES, Pedro Augusto et al. Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern. Frontiers in Microbiology, v. 12, Article 713713, p. 1-20, 2021.1664-302Xhttps://www.arca.fiocruz.br/handle/icict/5170510.3389/fmicb.2021.713713engFrontiers Research FoundationOptimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concerninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Microbiologia. Recife, PE, Brasil.Universidade Federal de Minas Gerais. Departamento de Física. Belo Horizonte, MG, Brasil.Fundação Hospitalar do Estado de Minas Gerais. Núcleo de Inovação Tecnológica. Belo Horizonte, MG, Brasil.Visuri Equipamentos e Serviços. Belo Horizonte, MG, Brasil / Universidade Federal de Minas Gerais. Departamento de Engenharia Elétrica. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Departamento de Física. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, BrasilFundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia e Núcleo de Bioinformática. Recife, PE, Brasil.Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.The coronavirus disease 2019 (COVID-19) pandemic unfolded due to the widespread severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission reinforced the urgent need for affordable molecular diagnostic alternative methods for massive testing screening. We present the clinical validation of a pH-dependent colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) for SARS-CoV-2 detection. The method revealed a limit of detection of 19.3 ± 2.7 viral genomic copies/μL when using RNA extracted samples obtained from nasopharyngeal swabs collected in guanidine-containing viral transport medium. Typical RT-LAMP reactions were performed at 65°C for 30 min. When compared to reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), up to cycle-threshold (Ct) value 32, RT-LAMP presented 98% [95% confidence interval (CI) = 95.3-99.5%] sensitivity and 100% (95% CI = 94.5-100%) specificity for SARS-CoV-2 RNA detection targeting E and N genes. No cross-reactivity was detected when testing other non-SARS-CoV virus, confirming high specificity. The test is compatible with primary RNA extraction-free samples. We also demonstrated that colorimetric RT-LAMP can detect SARS-CoV-2 variants of concern and variants of interest, such as variants occurring in Brazil named gamma (P.1), zeta (P.2), delta (B.1.617.2), B.1.1.374, and B.1.1.371. The method meets point-of-care requirements and can be deployed in the field for high-throughput COVID-19 testing campaigns, especially in countries where COVID-19 testing efforts are far from ideal to tackle the pandemics. Although RT-qPCR is considered the gold standard for SARS-CoV-2 RNA detection, it requires expensive equipment, infrastructure, and highly trained personnel. In contrast, RT-LAMP emerges as an affordable, inexpensive, and simple alternative for SARS-CoV-2 molecular detection that can be applied to massive COVID-19 testing campaigns and save lives.COVID-19RT-LAMPSARS-CoV-2Diagnostic testMolecular testRespiratory virusinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83082https://www.arca.fiocruz.br/bitstream/icict/51705/1/license.txt9193a7c197bc67acd023525e72a03240MD51ORIGINALAlves_Pedro_etal_IRR_IAM_COVID-19_2021.pdfAlves_Pedro_etal_IRR_IAM_COVID-19_2021.pdfapplication/pdf4014365https://www.arca.fiocruz.br/bitstream/icict/51705/2/Alves_Pedro_etal_IRR_IAM_COVID-19_2021.pdf288600a8b353372b04248fa895d0a89fMD52icict/517052022-03-17 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| dc.title.pt_BR.fl_str_mv |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| title |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| spellingShingle |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern Alves, Pedro Augusto COVID-19 RT-LAMP SARS-CoV-2 Diagnostic test Molecular test Respiratory virus |
| title_short |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| title_full |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| title_fullStr |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| title_full_unstemmed |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| title_sort |
Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern |
| author |
Alves, Pedro Augusto |
| author_facet |
Alves, Pedro Augusto Oliveira, Ellen Gonçalves de Luiz, Ana Paula Moreira Franco Almeida, Letícia Trindade Gonçalves, Amanda Bonoto Borges, Iara Apolinário Rocha, Flávia de S Rocha, Raissa Prado Bezerra, Matheus Filgueira Miranda, Pâmella Capanema, Flávio Diniz Martins, Henrique Resende Weber, Gerald Teixeira, Santuza Maria Ribeiro Wallau, Gabriel Luz Monte Neto, Rubens Lima do |
| author_role |
author |
| author2 |
Oliveira, Ellen Gonçalves de Luiz, Ana Paula Moreira Franco Almeida, Letícia Trindade Gonçalves, Amanda Bonoto Borges, Iara Apolinário Rocha, Flávia de S Rocha, Raissa Prado Bezerra, Matheus Filgueira Miranda, Pâmella Capanema, Flávio Diniz Martins, Henrique Resende Weber, Gerald Teixeira, Santuza Maria Ribeiro Wallau, Gabriel Luz Monte Neto, Rubens Lima do |
| author2_role |
author author author author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Alves, Pedro Augusto Oliveira, Ellen Gonçalves de Luiz, Ana Paula Moreira Franco Almeida, Letícia Trindade Gonçalves, Amanda Bonoto Borges, Iara Apolinário Rocha, Flávia de S Rocha, Raissa Prado Bezerra, Matheus Filgueira Miranda, Pâmella Capanema, Flávio Diniz Martins, Henrique Resende Weber, Gerald Teixeira, Santuza Maria Ribeiro Wallau, Gabriel Luz Monte Neto, Rubens Lima do |
| dc.subject.en.pt_BR.fl_str_mv |
COVID-19 RT-LAMP SARS-CoV-2 Diagnostic test Molecular test Respiratory virus |
| topic |
COVID-19 RT-LAMP SARS-CoV-2 Diagnostic test Molecular test Respiratory virus |
| description |
Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil. |
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2021 |
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2021 |
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2022-03-16T19:30:39Z |
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2022-03-16T19:30:39Z |
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info:eu-repo/semantics/article |
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ALVES, Pedro Augusto et al. Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern. Frontiers in Microbiology, v. 12, Article 713713, p. 1-20, 2021. |
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https://www.arca.fiocruz.br/handle/icict/51705 |
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1664-302X |
| dc.identifier.doi.none.fl_str_mv |
10.3389/fmicb.2021.713713 |
| identifier_str_mv |
ALVES, Pedro Augusto et al. Optimization and Clinical Validation of Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification, a Fast, Highly Sensitive and Specific COVID-19 Molecular Diagnostic Tool That Is Robust to Detect SARS-CoV-2 Variants of Concern. Frontiers in Microbiology, v. 12, Article 713713, p. 1-20, 2021. 1664-302X 10.3389/fmicb.2021.713713 |
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Frontiers Research Foundation |
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Frontiers Research Foundation |
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