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Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA)
SARS-CoV-2, a positive-strand RNA virus has caused devastating effects. The standard method for COVID diagnosis is based on polymerase chain reaction (PCR). The method needs expensive reagents and equipment and well-trained personnel and takes a few hours to be completed. The search for faster solut...
Autores principales: | , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9740288/ https://www.ncbi.nlm.nih.gov/pubmed/36499587 http://dx.doi.org/10.3390/ijms232315258 |
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author | Aviñó, Anna Cuestas-Ayllón, Carlos Gutiérrez-Capitán, Manuel Vilaplana, Lluisa Grazu, Valeria Noé, Véronique Balada, Eva Baldi, Antonio Félix, Alex J. Aubets, Eva Valiuska, Simonas Domínguez, Arnau Gargallo, Raimundo Eritja, Ramon Marco, M.-Pilar Fernández-Sánchez, César Martínez de la Fuente, Jesús Ciudad, Carlos J. |
author_facet | Aviñó, Anna Cuestas-Ayllón, Carlos Gutiérrez-Capitán, Manuel Vilaplana, Lluisa Grazu, Valeria Noé, Véronique Balada, Eva Baldi, Antonio Félix, Alex J. Aubets, Eva Valiuska, Simonas Domínguez, Arnau Gargallo, Raimundo Eritja, Ramon Marco, M.-Pilar Fernández-Sánchez, César Martínez de la Fuente, Jesús Ciudad, Carlos J. |
author_sort | Aviñó, Anna |
collection | PubMed |
description | SARS-CoV-2, a positive-strand RNA virus has caused devastating effects. The standard method for COVID diagnosis is based on polymerase chain reaction (PCR). The method needs expensive reagents and equipment and well-trained personnel and takes a few hours to be completed. The search for faster solutions has led to the development of immunological assays based on antibodies that recognize the viral proteins that are faster and do not require any special equipment. Here, we explore an innovative analytical approach based on the sandwich oligonucleotide hybridization which can be adapted to several biosensing devices including thermal lateral flow and electrochemical devices, as well as fluorescent microarrays. Polypurine reverse-Hoogsteen hairpins (PPRHs) oligonucleotides that form high-affinity triplexes with the polypyrimidine target sequences are used for the efficient capture of the viral genome. Then, a second labeled oligonucleotide is used to detect the formation of a trimolecular complex in a similar way to antigen tests. The reached limit of detection is around 0.01 nM (a few femtomoles) without the use of any amplification steps. The triplex enhanced nucleic acid detection assay (TENADA) can be readily adapted for the detection of any pathogen requiring only the knowledge of the pathogen genome sequence. |
format | Online Article Text |
id | pubmed-9740288 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-97402882022-12-11 Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) Aviñó, Anna Cuestas-Ayllón, Carlos Gutiérrez-Capitán, Manuel Vilaplana, Lluisa Grazu, Valeria Noé, Véronique Balada, Eva Baldi, Antonio Félix, Alex J. Aubets, Eva Valiuska, Simonas Domínguez, Arnau Gargallo, Raimundo Eritja, Ramon Marco, M.-Pilar Fernández-Sánchez, César Martínez de la Fuente, Jesús Ciudad, Carlos J. Int J Mol Sci Article SARS-CoV-2, a positive-strand RNA virus has caused devastating effects. The standard method for COVID diagnosis is based on polymerase chain reaction (PCR). The method needs expensive reagents and equipment and well-trained personnel and takes a few hours to be completed. The search for faster solutions has led to the development of immunological assays based on antibodies that recognize the viral proteins that are faster and do not require any special equipment. Here, we explore an innovative analytical approach based on the sandwich oligonucleotide hybridization which can be adapted to several biosensing devices including thermal lateral flow and electrochemical devices, as well as fluorescent microarrays. Polypurine reverse-Hoogsteen hairpins (PPRHs) oligonucleotides that form high-affinity triplexes with the polypyrimidine target sequences are used for the efficient capture of the viral genome. Then, a second labeled oligonucleotide is used to detect the formation of a trimolecular complex in a similar way to antigen tests. The reached limit of detection is around 0.01 nM (a few femtomoles) without the use of any amplification steps. The triplex enhanced nucleic acid detection assay (TENADA) can be readily adapted for the detection of any pathogen requiring only the knowledge of the pathogen genome sequence. MDPI 2022-12-03 /pmc/articles/PMC9740288/ /pubmed/36499587 http://dx.doi.org/10.3390/ijms232315258 Text en © 2022 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 Aviñó, Anna Cuestas-Ayllón, Carlos Gutiérrez-Capitán, Manuel Vilaplana, Lluisa Grazu, Valeria Noé, Véronique Balada, Eva Baldi, Antonio Félix, Alex J. Aubets, Eva Valiuska, Simonas Domínguez, Arnau Gargallo, Raimundo Eritja, Ramon Marco, M.-Pilar Fernández-Sánchez, César Martínez de la Fuente, Jesús Ciudad, Carlos J. Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title | Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title_full | Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title_fullStr | Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title_full_unstemmed | Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title_short | Detection of SARS-CoV-2 Virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA) |
title_sort | detection of sars-cov-2 virus by triplex enhanced nucleic acid detection assay (tenada) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9740288/ https://www.ncbi.nlm.nih.gov/pubmed/36499587 http://dx.doi.org/10.3390/ijms232315258 |
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