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

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dc.creatorPedro Augusto Alves
dc.creatorAna Paula Franco Luiz
dc.creatorLetícia Almeida
dc.creatorAmanda Gonçalves
dc.creatorFlávio Capanema
dc.creatorHenrique Resende Martins
dc.creatorGabriel Luz Wallau
dc.creatorRubens Monte Neto
dc.date.accessioned2023-09-14T19:25:51Z
dc.date.accessioned2025-09-09T00:58:40Z
dc.date.available2023-09-14T19:25:51Z
dc.date.issued2021
dc.identifier.doihttps://doi.org/10.3389/fmicb.2021.713713
dc.identifier.issn1664-302X
dc.identifier.urihttps://hdl.handle.net/1843/58693
dc.languagepor
dc.publisherUniversidade Federal de Minas Gerais
dc.relation.ispartofFrontiers in Microbiology
dc.rightsAcesso Aberto
dc.subjectCOVID-19
dc.subjectVírus sinciciais respiratórios
dc.subject.otherCOVID - 19
dc.subject.otherRT-LAMP
dc.subject.otherSARS-CoV-2
dc.subject.otherMolecular test
dc.subject.otherRespiratory virus
dc.subject.otherDiagnostic Test
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.typeArtigo de periódico
local.citation.epage20
local.citation.spage1
local.citation.volume12
local.description.resumoThe 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.
local.identifier.orcidhttps://orcid.org/0000-0002-4879-1345
local.identifier.orcidhttps://orcid.org/0000-0002-1419-5713
local.identifier.orcidhttps://orcid.org/0000-0002-4688-2462
local.publisher.countryBrasil
local.publisher.departmentENG - DEPARTAMENTO DE ENGENHARIA ELÉTRICA
local.publisher.departmentENG - DEPARTAMENTO DE ENGENHARIA ELETRÔNICA
local.publisher.departmentICB - DEPARTAMENTO DE BIOQUÍMICA E IMUNOLOGIA
local.publisher.departmentICX - DEPARTAMENTO DE FÍSICA
local.publisher.initialsUFMG
local.url.externahttps://www.frontiersin.org/articles/10.3389/fmicb.2021.713713/full

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