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Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples
The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-apt...
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/PMC9689637/ https://www.ncbi.nlm.nih.gov/pubmed/36428893 http://dx.doi.org/10.3390/diagnostics12112829 |
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author | Franco-Urquijo, Pablo Alberto Sierra-Martínez, Mónica Jarquín-Martínez, Mariana Martínez-Roque, Mateo Alejandro García-Velásquez, Victor Miguel Acosta-Altamirano, Gustavo Ruiz-Pérez, Nancy Jannet Toscano-Garibay, Julia Dolores Alvarez-Salas, Luis Marat |
author_facet | Franco-Urquijo, Pablo Alberto Sierra-Martínez, Mónica Jarquín-Martínez, Mariana Martínez-Roque, Mateo Alejandro García-Velásquez, Victor Miguel Acosta-Altamirano, Gustavo Ruiz-Pérez, Nancy Jannet Toscano-Garibay, Julia Dolores Alvarez-Salas, Luis Marat |
author_sort | Franco-Urquijo, Pablo Alberto |
collection | PubMed |
description | The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-aptamers (C7/C9) binding to the SARS-CoV-2 Spike (S) protein were isolated but its performance in clinical samples remained to be tested. Here, 242 samples were collected through three different methods and subjected to florescence-linked aptamer assays (FLAA) based on C7/C9 aptamers through two readout protocols. Then, a step-by-step statistical approach which included agreement tests, proportion comparisons and binomial and multinomial logistic regressions was used to predict optimal conditions for the novel C7/C9 FLAA test. RTqPCR threshold cycles, symptoms onset and processing time were influential factors on FLAA test results. Naturally occurring mutations on S were also detected and analyzed. Aminoacidic substitutions D614G and T732A appeared relevant for aptamer recognition although further studies are necessary. The methodology presented here is the first step to determine the performance and diagnosis across a range of clinical contexts and it might serve as a base for a complete analysis applicable to other designs of new diagnostic tests. |
format | Online Article Text |
id | pubmed-9689637 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-96896372022-11-25 Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples Franco-Urquijo, Pablo Alberto Sierra-Martínez, Mónica Jarquín-Martínez, Mariana Martínez-Roque, Mateo Alejandro García-Velásquez, Victor Miguel Acosta-Altamirano, Gustavo Ruiz-Pérez, Nancy Jannet Toscano-Garibay, Julia Dolores Alvarez-Salas, Luis Marat Diagnostics (Basel) Article The COVID-19 pandemic has been a main concern over the last two years and has become one of the most important crises in the history of human health. Today, there is still a need for affordable and reliable diagnostic tests for massive disease monitoring. Previously, a set of highly specific DNA-aptamers (C7/C9) binding to the SARS-CoV-2 Spike (S) protein were isolated but its performance in clinical samples remained to be tested. Here, 242 samples were collected through three different methods and subjected to florescence-linked aptamer assays (FLAA) based on C7/C9 aptamers through two readout protocols. Then, a step-by-step statistical approach which included agreement tests, proportion comparisons and binomial and multinomial logistic regressions was used to predict optimal conditions for the novel C7/C9 FLAA test. RTqPCR threshold cycles, symptoms onset and processing time were influential factors on FLAA test results. Naturally occurring mutations on S were also detected and analyzed. Aminoacidic substitutions D614G and T732A appeared relevant for aptamer recognition although further studies are necessary. The methodology presented here is the first step to determine the performance and diagnosis across a range of clinical contexts and it might serve as a base for a complete analysis applicable to other designs of new diagnostic tests. MDPI 2022-11-17 /pmc/articles/PMC9689637/ /pubmed/36428893 http://dx.doi.org/10.3390/diagnostics12112829 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 Franco-Urquijo, Pablo Alberto Sierra-Martínez, Mónica Jarquín-Martínez, Mariana Martínez-Roque, Mateo Alejandro García-Velásquez, Victor Miguel Acosta-Altamirano, Gustavo Ruiz-Pérez, Nancy Jannet Toscano-Garibay, Julia Dolores Alvarez-Salas, Luis Marat Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title | Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title_full | Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title_fullStr | Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title_full_unstemmed | Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title_short | Fluorescence-Linked Aptamer Assay for SARS-CoV-2 Spike-Protein: A Step-by-Step Performance Analysis in Clinical Samples |
title_sort | fluorescence-linked aptamer assay for sars-cov-2 spike-protein: a step-by-step performance analysis in clinical samples |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9689637/ https://www.ncbi.nlm.nih.gov/pubmed/36428893 http://dx.doi.org/10.3390/diagnostics12112829 |
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