Cargando…

Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification

The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused more than 6 million deaths all over the world, demonstrating the need for a simple, fast and cost-effective point-of-care (POC) test for the detection of the virus. In this work, we developed an electroche...

Descripción completa

Detalles Bibliográficos
Autores principales: Rivas-Macho, Ane, Eletxigerra, Unai, Diez-Ahedo, Ruth, Barros, Ángela, Merino, Santos, Goñi-de-Cerio, Felipe, Olabarria, Garbiñe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10605850/
https://www.ncbi.nlm.nih.gov/pubmed/37887117
http://dx.doi.org/10.3390/bios13100924
_version_ 1785127177289728000
author Rivas-Macho, Ane
Eletxigerra, Unai
Diez-Ahedo, Ruth
Barros, Ángela
Merino, Santos
Goñi-de-Cerio, Felipe
Olabarria, Garbiñe
author_facet Rivas-Macho, Ane
Eletxigerra, Unai
Diez-Ahedo, Ruth
Barros, Ángela
Merino, Santos
Goñi-de-Cerio, Felipe
Olabarria, Garbiñe
author_sort Rivas-Macho, Ane
collection PubMed
description The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused more than 6 million deaths all over the world, demonstrating the need for a simple, fast and cost-effective point-of-care (POC) test for the detection of the virus. In this work, we developed an electrochemical sensor for SARS-CoV-2 virus detection on clinical samples based on loop-mediated isothermal amplification (LAMP). With the development of this novel sensor, the time of each measurement is significantly reduced by avoiding the DNA extraction step and replacing it with inactivation of the sample by heating it at 95 °C for 10 min. To make the reaction compatible with the sample pre-treatment, an RNase inhibitor was added directly to the premix. The LAMP product was measured in a novel, easy-to-use manufactured sensor containing a custom-made screen-printed carbon electrode. Electrochemical detection was performed with a portable potentiostat, and methylene blue was used as the redox-transducing molecule. The developed sensor achieved a limit of detection of 62 viral copies and was 100% specific for the detection of the SARS-CoV-2 virus. The performance of the electrochemical sensor was validated with nasopharyngeal samples, obtaining a sensibility and specificity of 100% compared to the gold standard RT-PCR method.
format Online
Article
Text
id pubmed-10605850
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-106058502023-10-28 Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification Rivas-Macho, Ane Eletxigerra, Unai Diez-Ahedo, Ruth Barros, Ángela Merino, Santos Goñi-de-Cerio, Felipe Olabarria, Garbiñe Biosensors (Basel) Article The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused more than 6 million deaths all over the world, demonstrating the need for a simple, fast and cost-effective point-of-care (POC) test for the detection of the virus. In this work, we developed an electrochemical sensor for SARS-CoV-2 virus detection on clinical samples based on loop-mediated isothermal amplification (LAMP). With the development of this novel sensor, the time of each measurement is significantly reduced by avoiding the DNA extraction step and replacing it with inactivation of the sample by heating it at 95 °C for 10 min. To make the reaction compatible with the sample pre-treatment, an RNase inhibitor was added directly to the premix. The LAMP product was measured in a novel, easy-to-use manufactured sensor containing a custom-made screen-printed carbon electrode. Electrochemical detection was performed with a portable potentiostat, and methylene blue was used as the redox-transducing molecule. The developed sensor achieved a limit of detection of 62 viral copies and was 100% specific for the detection of the SARS-CoV-2 virus. The performance of the electrochemical sensor was validated with nasopharyngeal samples, obtaining a sensibility and specificity of 100% compared to the gold standard RT-PCR method. MDPI 2023-10-11 /pmc/articles/PMC10605850/ /pubmed/37887117 http://dx.doi.org/10.3390/bios13100924 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Rivas-Macho, Ane
Eletxigerra, Unai
Diez-Ahedo, Ruth
Barros, Ángela
Merino, Santos
Goñi-de-Cerio, Felipe
Olabarria, Garbiñe
Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title_full Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title_fullStr Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title_full_unstemmed Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title_short Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification
title_sort development of an electrochemical sensor for sars-cov-2 detection based on loop-mediated isothermal amplification
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10605850/
https://www.ncbi.nlm.nih.gov/pubmed/37887117
http://dx.doi.org/10.3390/bios13100924
work_keys_str_mv AT rivasmachoane developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT eletxigerraunai developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT diezahedoruth developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT barrosangela developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT merinosantos developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT gonideceriofelipe developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification
AT olabarriagarbine developmentofanelectrochemicalsensorforsarscov2detectionbasedonloopmediatedisothermalamplification