Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system

Detalhes bibliográficos
Autor(a) principal: Sousa, Karoline Almeida Felix de
Data de Publicação: 2023
Outros Autores: Nonaka, Carolina Kymie Vasques, Khouri, Ricardo, Rocha, Clarissa Araújo Gurgel, Silva, Carlos Gustavo Regis, Souza, Bruno Solano de Freitas
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/60838
Resumo: Inova Fiocruz. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
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spelling Sousa, Karoline Almeida Felix deNonaka, Carolina Kymie VasquesKhouri, RicardoRocha, Clarissa Araújo GurgelSilva, Carlos Gustavo RegisSouza, Bruno Solano de Freitas2023-10-18T17:58:54Z2023-10-18T17:58:54Z2023SOUSA, Karoline Almeida Felix de et al. Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system. Diagnostics, v.13, n. 13, p. 1-7, 2023.2075-4418https://www.arca.fiocruz.br/handle/icict/6083810.3390/diagnostics13132233Inova Fiocruz. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Instituto D’Or de Pesquisa e Ensino (IDOR). Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil.Instituto D’Or de Pesquisa e Ensino (IDOR). Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Hospital São Rafael. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Instituto D’Or de Pesquisa e Ensino (IDOR). Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Odontologia. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil.Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Instituto D’Or de Pesquisa e Ensino (IDOR). Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Hospital São Rafael. Salvador, BA, Brasil.Background: The global public health system has been severely tested by the COVID-19 pandemic. Mass testing was essential in controlling the transmission of the SARS-CoV-2; however, its implementation has encountered challenges, particularly in low-income countries. The urgent need for rapid and accurate tests for SARS-CoV-2 has proven to be extremely important. Pointof-care tests using the CRISPR system for COVID-19 have shown promise, with a reported high sensitivity and rapid detection. The performance of a CRISPR-based SARS-CoV-2 testing system was reported in this study. Methods: A total of 29 nasopharyngeal samples were evaluated, including 23 samples from individuals suspected of COVID-19, and six samples positive for H3N2 or respiratory syncytial virus. Two reference samples with known concentrations of SARS-CoV-2 RNA (3000 RNA copies/mL) or viral titer determined by plaque assay (105 PFU/mL) were also evaluated. The LAMP technique was employed to amplify the ORF1ab gene and the results were analyzed using a Gemini XPS fluorescence reader. Results: The RT-LAMP-CRISPR/Cas12 assay showed 100% concordance compared to RT-PCR. The RT-PCR presented a detection limit of 0.01 PFU/mL and the CRISPR/Cas12 system showed a limit of 15.6 PFU/mL. The RT-PCR sensitivity was approximately 8 RNA copies/ L and CRISPR/Cas12 at 84 RNA copies/ L. Conclusion: The RT-LAMP-CRISPR/Cas12a assay offered a promising alternative for the detection of SARS-CoV-2 and reinforces that CRISPR-based diagnostic techniques can be an alternative for fast and accurate assays.engMDPICOVID 19Ponto de atendimentoCRISPRControle de infecçãoDiagnósticoCOVID-19Point-of-careCRISPRInfection controlDiagnostic testCOVID 19Repetições Palindrômicas Curtas Agrupadas e Regularmente EspaçadasControle de InfecçõesTécnicas e Procedimentos DiagnósticosRapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a systeminfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALSousa, K. A. F. - Rapid Detection of SARS-CoV-2 Based on the LAMP Assay Associated with the CRISPRCas12a System.pdfSousa, K. A. F. - Rapid Detection of SARS-CoV-2 Based on the LAMP Assay Associated with the CRISPRCas12a System.pdfapplication/pdf1557527https://www.arca.fiocruz.br/bitstream/icict/60838/2/Sousa%2c%20K.%20A.%20F.%20-%20Rapid%20Detection%20of%20SARS-CoV-2%20Based%20on%20the%20LAMP%20Assay%20Associated%20with%20the%20CRISPRCas12a%20System.pdf1f45530d74049199d94a0cfc0ef449dbMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/60838/1/license.txt5a560609d32a3863062d77ff32785d58MD51icict/608382023-10-18 14:58:55.346oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352023-10-18T17:58:55Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.en_US.fl_str_mv Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
title Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
spellingShingle Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
Sousa, Karoline Almeida Felix de
COVID 19
Ponto de atendimento
CRISPR
Controle de infecção
Diagnóstico
COVID-19
Point-of-care
CRISPR
Infection control
Diagnostic test
COVID 19
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
Controle de Infecções
Técnicas e Procedimentos Diagnósticos
title_short Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
title_full Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
title_fullStr Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
title_full_unstemmed Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
title_sort Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system
author Sousa, Karoline Almeida Felix de
author_facet Sousa, Karoline Almeida Felix de
Nonaka, Carolina Kymie Vasques
Khouri, Ricardo
Rocha, Clarissa Araújo Gurgel
Silva, Carlos Gustavo Regis
Souza, Bruno Solano de Freitas
author_role author
author2 Nonaka, Carolina Kymie Vasques
Khouri, Ricardo
Rocha, Clarissa Araújo Gurgel
Silva, Carlos Gustavo Regis
Souza, Bruno Solano de Freitas
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Sousa, Karoline Almeida Felix de
Nonaka, Carolina Kymie Vasques
Khouri, Ricardo
Rocha, Clarissa Araújo Gurgel
Silva, Carlos Gustavo Regis
Souza, Bruno Solano de Freitas
dc.subject.other.en_US.fl_str_mv COVID 19
Ponto de atendimento
CRISPR
Controle de infecção
Diagnóstico
topic COVID 19
Ponto de atendimento
CRISPR
Controle de infecção
Diagnóstico
COVID-19
Point-of-care
CRISPR
Infection control
Diagnostic test
COVID 19
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
Controle de Infecções
Técnicas e Procedimentos Diagnósticos
dc.subject.en.en_US.fl_str_mv COVID-19
Point-of-care
CRISPR
Infection control
Diagnostic test
dc.subject.decs.en_US.fl_str_mv COVID 19
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
Controle de Infecções
Técnicas e Procedimentos Diagnósticos
description Inova Fiocruz. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
publishDate 2023
dc.date.accessioned.fl_str_mv 2023-10-18T17:58:54Z
dc.date.available.fl_str_mv 2023-10-18T17:58:54Z
dc.date.issued.fl_str_mv 2023
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv SOUSA, Karoline Almeida Felix de et al. Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system. Diagnostics, v.13, n. 13, p. 1-7, 2023.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/60838
dc.identifier.issn.en_US.fl_str_mv 2075-4418
dc.identifier.doi.none.fl_str_mv 10.3390/diagnostics13132233
identifier_str_mv SOUSA, Karoline Almeida Felix de et al. Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system. Diagnostics, v.13, n. 13, p. 1-7, 2023.
2075-4418
10.3390/diagnostics13132233
url https://www.arca.fiocruz.br/handle/icict/60838
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
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