Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update
The emergence of the COVID-19 pandemic prompted fast development of novel diagnostic methods of the etiologic virus SARS-CoV-2. Methods based on CRISPR–Cas systems have been particularly promising because they can achieve a similar sensitivity and specificity to the benchmark RT-qPCR, especially whe...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/12/6/1434 |
| _version_ | 1797488256164560896 |
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| author | Armando Hernandez-Garcia Melissa D. Morales-Moreno Erick G. Valdés-Galindo Eric P. Jimenez-Nieto Andrea Quezada |
| author_facet | Armando Hernandez-Garcia Melissa D. Morales-Moreno Erick G. Valdés-Galindo Eric P. Jimenez-Nieto Andrea Quezada |
| author_sort | Armando Hernandez-Garcia |
| collection | DOAJ |
| description | The emergence of the COVID-19 pandemic prompted fast development of novel diagnostic methods of the etiologic virus SARS-CoV-2. Methods based on CRISPR–Cas systems have been particularly promising because they can achieve a similar sensitivity and specificity to the benchmark RT-qPCR, especially when coupled to an isothermal pre-amplification step. Furthermore, they have also solved inherent limitations of RT-qPCR that impede its decentralized use and deployment in the field, such as the need for expensive equipment, high cost per reaction, and delivery of results in hours, among others. In this review, we evaluate publicly available methods to detect SARS-CoV-2 that are based on CRISPR–Cas and isothermal amplification. We critically analyze the steps required to obtain a successful result from clinical samples and pinpoint key experimental conditions and parameters that could be optimized or modified to improve clinical and analytical outputs. The COVID outbreak has propelled intensive research in a short time, which is paving the way to develop effective and very promising CRISPR–Cas systems for the precise detection of SARS-CoV-2. This review could also serve as an introductory guide to new labs delving into this technology. |
| first_indexed | 2024-03-10T00:00:31Z |
| format | Article |
| id | doaj.art-8ca43690a28842a88e740fda7754b5fa |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-10T00:00:31Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-8ca43690a28842a88e740fda7754b5fa2023-11-23T16:18:06ZengMDPI AGDiagnostics2075-44182022-06-01126143410.3390/diagnostics12061434Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and UpdateArmando Hernandez-Garcia0Melissa D. Morales-Moreno1Erick G. Valdés-Galindo2Eric P. Jimenez-Nieto3Andrea Quezada4Laboratory of Biomolecular Engineering and Bionanotechnology, Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, Ciudad de Mexico C.P. 04510, MexicoLaboratory of Biomolecular Engineering and Bionanotechnology, Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, Ciudad de Mexico C.P. 04510, MexicoLaboratory of Biomolecular Engineering and Bionanotechnology, Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, Ciudad de Mexico C.P. 04510, MexicoLaboratory of Biomolecular Engineering and Bionanotechnology, Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, Ciudad de Mexico C.P. 04510, MexicoLaboratory of Biomolecular Engineering and Bionanotechnology, Department of Chemistry of Biomacromolecules, Institute of Chemistry, National Autonomous University of Mexico, Circuito Exterior, Ciudad Universitaria, Coyoacan, Ciudad de Mexico C.P. 04510, MexicoThe emergence of the COVID-19 pandemic prompted fast development of novel diagnostic methods of the etiologic virus SARS-CoV-2. Methods based on CRISPR–Cas systems have been particularly promising because they can achieve a similar sensitivity and specificity to the benchmark RT-qPCR, especially when coupled to an isothermal pre-amplification step. Furthermore, they have also solved inherent limitations of RT-qPCR that impede its decentralized use and deployment in the field, such as the need for expensive equipment, high cost per reaction, and delivery of results in hours, among others. In this review, we evaluate publicly available methods to detect SARS-CoV-2 that are based on CRISPR–Cas and isothermal amplification. We critically analyze the steps required to obtain a successful result from clinical samples and pinpoint key experimental conditions and parameters that could be optimized or modified to improve clinical and analytical outputs. The COVID outbreak has propelled intensive research in a short time, which is paving the way to develop effective and very promising CRISPR–Cas systems for the precise detection of SARS-CoV-2. This review could also serve as an introductory guide to new labs delving into this technology.https://www.mdpi.com/2075-4418/12/6/1434CRISPR–CasSARS-CoV-2molecular diagnosticsisothermal amplificationcomparative analysisnucleic acid detection |
| spellingShingle | Armando Hernandez-Garcia Melissa D. Morales-Moreno Erick G. Valdés-Galindo Eric P. Jimenez-Nieto Andrea Quezada Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update Diagnostics CRISPR–Cas SARS-CoV-2 molecular diagnostics isothermal amplification comparative analysis nucleic acid detection |
| title | Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update |
| title_full | Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update |
| title_fullStr | Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update |
| title_full_unstemmed | Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update |
| title_short | Diagnostics of COVID-19 Based on CRISPR–Cas Coupled to Isothermal Amplification: A Comparative Analysis and Update |
| title_sort | diagnostics of covid 19 based on crispr cas coupled to isothermal amplification a comparative analysis and update |
| topic | CRISPR–Cas SARS-CoV-2 molecular diagnostics isothermal amplification comparative analysis nucleic acid detection |
| url | https://www.mdpi.com/2075-4418/12/6/1434 |
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