Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification

Evaluation of the performance of a new set of primers defined from the ORF1ab sequence, and its combination with a previously published set of primers from the N sequence, to detect SARS-CoV-2 RNA by the loop-mediated isothermal amplification technique is presented. The ORF1ab primer set enables vis...

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Authors: Reynés, Bàrbara, Serra, Francisca, Palou, Andreu
Format: article
Publication Date:2021
Country:España
Institution:Conselleria de Salut i Consum del Govern de les Illes Balears
Repository:Docusalut
Language:English
OAI Identifier:oai:docusalut.com:20.500.13003/19789
Online Access:https://hdl.handle.net/20.500.13003/19789
Access Level:Open access
Keyword:Nucleic Acid Amplification Techniques
Polyproteins
SARS-CoV-2
Colorimetry
DNA Primers
Humans
COVID-19
Molecular Diagnostic Techniques
Viral Proteins
COVID-19 Nucleic Acid Testing
Prueba de Ácido Nucleico para COVID-19
Colorimetría
Humanos
Proteínas Virales
Cartilla de ADN
Técnicas de Diagnóstico Molecular
Poliproteínas
Técnicas de Amplificación de Ácido Nucleico
nucleocapsid primers
ORF1ab primers
SARS-CoV-2 detection
virus LAMP amplification
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spelling Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplificationReynés, BàrbaraSerra, FranciscaPalou, AndreuNucleic Acid Amplification TechniquesPolyproteinsSARS-CoV-2ColorimetryDNA PrimersHumansCOVID-19Molecular Diagnostic TechniquesViral ProteinsCOVID-19 Nucleic Acid TestingPrueba de Ácido Nucleico para COVID-19ColorimetríaHumanosSARS-CoV-2Proteínas ViralesCartilla de ADNTécnicas de Diagnóstico MolecularCOVID-19PoliproteínasTécnicas de Amplificación de Ácido NucleicoCOVID-19nucleocapsid primersORF1ab primersSARS-CoV-2 detectionvirus LAMP amplificationEvaluation of the performance of a new set of primers defined from the ORF1ab sequence, and its combination with a previously published set of primers from the N sequence, to detect SARS-CoV-2 RNA by the loop-mediated isothermal amplification technique is presented. The ORF1ab primer set enables visual detection of SARS-CoV-2 RNA in 16 min. In addition, a simultaneous reaction with both ORF1ab and N primers allows for higher sensitivity of detection, particularly when low numbers of copies are present (250 viral RNA copies). Further, the protocol is able to detect viral RNA in saliva samples. The procedure reported could be easily implemented in the generation of a new and sensitive rapid point-of care device for SARS-CoV-2 RNA visual detection. METHOD SUMMARY Optimized loop-mediated isothermal amplification reactions were carried out using the combination of two sets of primers (designed on ORF1ab and N sequences). Saliva samples were pretreated by heating and proteinase K, before SARS-CoV-2 determination by loop-mediated isothermal amplification.Future Sci Ltd20212021-04-0120212021-04-01research articlehttp://purl.org/coar/resource_type/c_2df8fbb1info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/20.500.13003/19789reponame:Docusalutinstname:Conselleria de Salut i Consum del Govern de les Illes BalearsInglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution-NonCommercial-ShareAlike 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-sa/4.0/info:eu-repo/semantics/openAccessoai:docusalut.com:20.500.13003/197892026-06-22T12:44:07Z
dc.title.none.fl_str_mv Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
title Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
spellingShingle Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
Reynés, Bàrbara
Nucleic Acid Amplification Techniques
Polyproteins
SARS-CoV-2
Colorimetry
DNA Primers
Humans
COVID-19
Molecular Diagnostic Techniques
Viral Proteins
COVID-19 Nucleic Acid Testing
Prueba de Ácido Nucleico para COVID-19
Colorimetría
Humanos
SARS-CoV-2
Proteínas Virales
Cartilla de ADN
Técnicas de Diagnóstico Molecular
COVID-19
Poliproteínas
Técnicas de Amplificación de Ácido Nucleico
COVID-19
nucleocapsid primers
ORF1ab primers
SARS-CoV-2 detection
virus LAMP amplification
title_short Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
title_full Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
title_fullStr Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
title_full_unstemmed Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
title_sort Rapid visual detection of SARS-CoV-2 by colorimetric loop-mediated isothermal amplification
dc.creator.none.fl_str_mv Reynés, Bàrbara
Serra, Francisca
Palou, Andreu
author Reynés, Bàrbara
author_facet Reynés, Bàrbara
Serra, Francisca
Palou, Andreu
author_role author
author2 Serra, Francisca
Palou, Andreu
author2_role author
author
dc.contributor.none.fl_str_mv
dc.subject.none.fl_str_mv Nucleic Acid Amplification Techniques
Polyproteins
SARS-CoV-2
Colorimetry
DNA Primers
Humans
COVID-19
Molecular Diagnostic Techniques
Viral Proteins
COVID-19 Nucleic Acid Testing
Prueba de Ácido Nucleico para COVID-19
Colorimetría
Humanos
SARS-CoV-2
Proteínas Virales
Cartilla de ADN
Técnicas de Diagnóstico Molecular
COVID-19
Poliproteínas
Técnicas de Amplificación de Ácido Nucleico
COVID-19
nucleocapsid primers
ORF1ab primers
SARS-CoV-2 detection
virus LAMP amplification
topic Nucleic Acid Amplification Techniques
Polyproteins
SARS-CoV-2
Colorimetry
DNA Primers
Humans
COVID-19
Molecular Diagnostic Techniques
Viral Proteins
COVID-19 Nucleic Acid Testing
Prueba de Ácido Nucleico para COVID-19
Colorimetría
Humanos
SARS-CoV-2
Proteínas Virales
Cartilla de ADN
Técnicas de Diagnóstico Molecular
COVID-19
Poliproteínas
Técnicas de Amplificación de Ácido Nucleico
COVID-19
nucleocapsid primers
ORF1ab primers
SARS-CoV-2 detection
virus LAMP amplification
description Evaluation of the performance of a new set of primers defined from the ORF1ab sequence, and its combination with a previously published set of primers from the N sequence, to detect SARS-CoV-2 RNA by the loop-mediated isothermal amplification technique is presented. The ORF1ab primer set enables visual detection of SARS-CoV-2 RNA in 16 min. In addition, a simultaneous reaction with both ORF1ab and N primers allows for higher sensitivity of detection, particularly when low numbers of copies are present (250 viral RNA copies). Further, the protocol is able to detect viral RNA in saliva samples. The procedure reported could be easily implemented in the generation of a new and sensitive rapid point-of care device for SARS-CoV-2 RNA visual detection. METHOD SUMMARY Optimized loop-mediated isothermal amplification reactions were carried out using the combination of two sets of primers (designed on ORF1ab and N sequences). Saliva samples were pretreated by heating and proteinase K, before SARS-CoV-2 determination by loop-mediated isothermal amplification.
publishDate 2021
dc.date.none.fl_str_mv 2021
2021-04-01
2021
2021-04-01
dc.type.none.fl_str_mv research article
http://purl.org/coar/resource_type/c_2df8fbb1
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/20.500.13003/19789
url https://hdl.handle.net/20.500.13003/19789
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution-NonCommercial-ShareAlike 4.0 International
http://creativecommons.org/licenses/by-nc-sa/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Future Sci Ltd
publisher.none.fl_str_mv Future Sci Ltd
dc.source.none.fl_str_mv reponame:Docusalut
instname:Conselleria de Salut i Consum del Govern de les Illes Balears
instname_str Conselleria de Salut i Consum del Govern de les Illes Balears
reponame_str Docusalut
collection Docusalut
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,812429

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