| Author | Alves, Pedro Augusto | |
| Author | Oliveira, Ellen Gonçalves de | |
| Author | Luiz, Ana Paula Moreira Franco | |
| Author | Almeida, Letícia Trindade | |
| Author | Gonçalves, Amanda Bonoto | |
| Author | Borges, Iara Apolinário | |
| Author | Rocha, Flávia de S | |
| Author | Rocha, Raissa Prado | |
| Author | Bezerra, Matheus Filgueira | |
| Author | Miranda, Pâmella | |
| Author | Capanema, Flávio Diniz | |
| Author | Martins, Henrique Resende | |
| Author | Weber, Gerald | |
| Author | Teixeira, Santuza Maria Ribeiro | |
| Author | Wallau, Gabriel Luz | |
| Author | Monte Neto, Rubens Lima do | |
| Access date | 2022-03-16T19:30:39Z | |
| Available date | 2022-03-16T19:30:39Z | |
| Document date | 2021 | |
| Citation | 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. | pt_BR |
| ISSN | 1664-302X | pt_BR |
| URI | https://www.arca.fiocruz.br/handle/icict/51705 | |
| Language | eng | pt_BR |
| Publisher | Frontiers Research Foundation | pt_BR |
| Rights | open access | pt_BR |
| 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 | pt_BR |
| Type | Article | pt_BR |
| DOI | 10.3389/fmicb.2021.713713 | |
| Abstract | 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. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/ Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Microbiologia. Recife, PE, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Minas Gerais. Departamento de Física. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Fundação Hospitalar do Estado de Minas Gerais. Núcleo de Inovação Tecnológica. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Visuri Equipamentos e Serviços. Belo Horizonte, MG, Brasil / Universidade Federal de Minas Gerais. Departamento de Engenharia Elétrica. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Minas Gerais. Departamento de Física. Belo Horizonte, MG, Brasil. | pt_BR |
| Affilliation | Universidade Federal de Minas Gerais. Centro de Tecnologia em Vacinas. Belo Horizonte, MG, Brasil | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Aggeu Magalhães. Departamento de Entomologia e Núcleo de Bioinformática. Recife, PE, Brasil. | pt_BR |
| Affilliation | Fundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil. | pt_BR |
| Subject | COVID-19 | pt_BR |
| Subject | RT-LAMP | pt_BR |
| Subject | SARS-CoV-2 | pt_BR |
| Subject | Diagnostic test | pt_BR |
| Subject | Molecular test | pt_BR |
| Subject | Respiratory virus | pt_BR |
