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Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2

The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs...

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Detalles Bibliográficos
Autores principales: Bektaş, Ali, Covington, Michael F., Aidelberg, Guy, Arce, Anibal, Matute, Tamara, Núñez, Isaac, Walsh, Julia, Boutboul, David, Delaugerre, Constance, Lindner, Ariel B., Federici, Fernán, Jayaprakash, Anitha D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146324/
https://www.ncbi.nlm.nih.gov/pubmed/33922716
http://dx.doi.org/10.3390/v13050742
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author Bektaş, Ali
Covington, Michael F.
Aidelberg, Guy
Arce, Anibal
Matute, Tamara
Núñez, Isaac
Walsh, Julia
Boutboul, David
Delaugerre, Constance
Lindner, Ariel B.
Federici, Fernán
Jayaprakash, Anitha D.
author_facet Bektaş, Ali
Covington, Michael F.
Aidelberg, Guy
Arce, Anibal
Matute, Tamara
Núñez, Isaac
Walsh, Julia
Boutboul, David
Delaugerre, Constance
Lindner, Ariel B.
Federici, Fernán
Jayaprakash, Anitha D.
author_sort Bektaş, Ali
collection PubMed
description The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs and supply shortages. Antigen tests that bind and detect the surface proteins of a virus are rapid and scalable but suffer from high false negative rates. To address this problem, an inexpensive, simple, and robust 60-minute do-it-yourself (DIY) workflow to detect viral RNA from nasal swabs or saliva with high sensitivity (0.1 to 2 viral particles/μL) and specificity (>97% true negative rate) utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed. ALERT (Accessible LAMP-Enabled Rapid Test) incorporates the following features: (1) increased shelf-life and ambient temperature storage, compared to liquid reaction mixes, by using wax layers to isolate enzymes from other reagents; (2) improved specificity compared to other LAMP end-point reporting methods, by using sequence-specific QUASR (quenching of unincorporated amplification signal reporters); (3) increased sensitivity, compared to methods without purification through use of a magnetic wand to enable pipette-free concentration of sample RNA and cell debris removal; (4) quality control with a nasopharyngeal-specific mRNA target; and (5) co-detection of other respiratory viruses, such as influenza B, by multiplexing QUASR-modified RT-LAMP primer sets. The flexible nature of the ALERT workflow allows easy, at-home and point-of-care testing for individuals and higher-throughput processing for labs and hospitals. With minimal effort, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific primer sets can be swapped out for other targets to repurpose ALERT to detect other viruses, microorganisms, or nucleic acid-based markers.
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spelling pubmed-81463242021-05-26 Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2 Bektaş, Ali Covington, Michael F. Aidelberg, Guy Arce, Anibal Matute, Tamara Núñez, Isaac Walsh, Julia Boutboul, David Delaugerre, Constance Lindner, Ariel B. Federici, Fernán Jayaprakash, Anitha D. Viruses Article The coronavirus disease 2019 (COVID-19) pandemic has highlighted bottlenecks in large-scale, frequent testing of populations for infections. Polymerase chain reaction (PCR)-based diagnostic tests are expensive, reliant on centralized labs, can take days to deliver results, and are prone to backlogs and supply shortages. Antigen tests that bind and detect the surface proteins of a virus are rapid and scalable but suffer from high false negative rates. To address this problem, an inexpensive, simple, and robust 60-minute do-it-yourself (DIY) workflow to detect viral RNA from nasal swabs or saliva with high sensitivity (0.1 to 2 viral particles/μL) and specificity (>97% true negative rate) utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) was developed. ALERT (Accessible LAMP-Enabled Rapid Test) incorporates the following features: (1) increased shelf-life and ambient temperature storage, compared to liquid reaction mixes, by using wax layers to isolate enzymes from other reagents; (2) improved specificity compared to other LAMP end-point reporting methods, by using sequence-specific QUASR (quenching of unincorporated amplification signal reporters); (3) increased sensitivity, compared to methods without purification through use of a magnetic wand to enable pipette-free concentration of sample RNA and cell debris removal; (4) quality control with a nasopharyngeal-specific mRNA target; and (5) co-detection of other respiratory viruses, such as influenza B, by multiplexing QUASR-modified RT-LAMP primer sets. The flexible nature of the ALERT workflow allows easy, at-home and point-of-care testing for individuals and higher-throughput processing for labs and hospitals. With minimal effort, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-specific primer sets can be swapped out for other targets to repurpose ALERT to detect other viruses, microorganisms, or nucleic acid-based markers. MDPI 2021-04-23 /pmc/articles/PMC8146324/ /pubmed/33922716 http://dx.doi.org/10.3390/v13050742 Text en © 2021 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
Bektaş, Ali
Covington, Michael F.
Aidelberg, Guy
Arce, Anibal
Matute, Tamara
Núñez, Isaac
Walsh, Julia
Boutboul, David
Delaugerre, Constance
Lindner, Ariel B.
Federici, Fernán
Jayaprakash, Anitha D.
Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title_full Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title_fullStr Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title_full_unstemmed Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title_short Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2
title_sort accessible lamp-enabled rapid test (alert) for detecting sars-cov-2
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8146324/
https://www.ncbi.nlm.nih.gov/pubmed/33922716
http://dx.doi.org/10.3390/v13050742
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