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

dc.creatorAlves, Pedro Augusto
dc.creatorOliveira, Ellen Gonçalves de
dc.creatorLuiz, Ana Paula Moreira Franco
dc.creatorAlmeida, Letícia Trindade
dc.creatorGonçalves, Amanda Bonoto
dc.creatorBorges, Iara Apolinário
dc.creatorRocha, Flávia de S
dc.creatorRocha, Raissa Prado
dc.creatorBezerra, Matheus Filgueira
dc.creatorMiranda, Pâmella
dc.creatorCapanema, Flávio Diniz
dc.creatorMartins, Henrique Resende
dc.creatorWeber, Gerald
dc.creatorTeixeira, Santuza Maria Ribeiro
dc.creatorWallau, Gabriel Luz
dc.creatorMonte Neto, Rubens Lima do
dc.date2022-03-16T19:30:39Z
dc.date2022-03-16T19:30:39Z
dc.date2021
dc.date.accessioned2023-09-26T23:21:39Z
dc.date.available2023-09-26T23:21:39Z
dc.identifierALVES, 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.
dc.identifier1664-302X
dc.identifierhttps://www.arca.fiocruz.br/handle/icict/51705
dc.identifier10.3389/fmicb.2021.713713
dc.identifier.urihttps://repositorioslatinoamericanos.uchile.cl/handle/2250/8889726
dc.descriptionThe 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.
dc.formatapplication/pdf
dc.languageeng
dc.publisherFrontiers Research Foundation
dc.rightsopen access
dc.subjectCOVID-19
dc.subjectRT-LAMP
dc.subjectSARS-CoV-2
dc.subjectDiagnostic test
dc.subjectMolecular test
dc.subjectRespiratory virus
dc.titleOptimization 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
dc.typeArticle


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