Author | Sousa, Karoline Almeida Felix de | |
Author | Nonaka, Carolina Kymie Vasques | |
Author | Khouri, Ricardo | |
Author | Rocha, Clarissa Araújo Gurgel | |
Author | Silva, Carlos Gustavo Regis | |
Author | Souza, Bruno Solano de Freitas | |
Access date | 2023-10-18T17:58:54Z | |
Available date | 2023-10-18T17:58:54Z | |
Document date | 2023 | |
Citation | 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. | en_US |
ISSN | 2075-4418 | en_US |
URI | https://www.arca.fiocruz.br/handle/icict/60838 | |
Sponsorship | Inova Fiocruz.
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). | en_US |
Language | eng | en_US |
Publisher | MDPI | en_US |
Rights | open access | en_US |
Subject in Portuguese | COVID 19 | en_US |
Subject in Portuguese | Ponto de atendimento | en_US |
Subject in Portuguese | CRISPR | en_US |
Subject in Portuguese | Controle de infecção | en_US |
Subject in Portuguese | Diagnóstico | en_US |
Title | Rapid detection of SARS-CoV-2 based on the LAMP assay associated with the CRISPRCas12a system | en_US |
Type | Article | en_US |
DOI | 10.3390/diagnostics13132233 | |
Abstract | 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. | en_US |
Affilliation | 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. | en_US |
Affilliation | Instituto D’Or de Pesquisa e Ensino (IDOR). Centro de Biotecnologia e Terapia Celular. Salvador, BA, Brasil / Hospital São Rafael. Salvador, BA, Brasil. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Universidade Federal da Bahia. Faculdade de Medicina. Salvador, BA, Brasil. | en_US |
Affilliation | 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. | en_US |
Affilliation | Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil. | en_US |
Affilliation | 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. | en_US |
Subject | COVID-19 | en_US |
Subject | Point-of-care | en_US |
Subject | CRISPR | en_US |
Subject | Infection control | en_US |
Subject | Diagnostic test | en_US |
DeCS | COVID 19 | en_US |
DeCS | Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas | en_US |
DeCS | Controle de Infecções | en_US |
DeCS | Técnicas e Procedimentos Diagnósticos | en_US |