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A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples

We report a small-footprint cost-effective isothermal rapid DNA amplification system, with integrated microfluidics for automated sample analysis and detection of SARS-CoV-2 in human and environmental samples. Our system measures low-level fluorescent signals in real-time during amplification, while...

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Autores principales: Staples, Jake, Dourou, Athanasia-Maria, Liampa, Irene, Sjaarda, Calvin, Moslinger, Emily, Wong, Henry, Sheth, Prameet M., Arhondakis, Stilianos, Prakash, Ravi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10377682/
https://www.ncbi.nlm.nih.gov/pubmed/37509680
http://dx.doi.org/10.3390/biomedicines11072038
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author Staples, Jake
Dourou, Athanasia-Maria
Liampa, Irene
Sjaarda, Calvin
Moslinger, Emily
Wong, Henry
Sheth, Prameet M.
Arhondakis, Stilianos
Prakash, Ravi
author_facet Staples, Jake
Dourou, Athanasia-Maria
Liampa, Irene
Sjaarda, Calvin
Moslinger, Emily
Wong, Henry
Sheth, Prameet M.
Arhondakis, Stilianos
Prakash, Ravi
author_sort Staples, Jake
collection PubMed
description We report a small-footprint cost-effective isothermal rapid DNA amplification system, with integrated microfluidics for automated sample analysis and detection of SARS-CoV-2 in human and environmental samples. Our system measures low-level fluorescent signals in real-time during amplification, while maintaining the desired assay temperature on a low power, portable system footprint. A unique soft microfluidic chip design was implemented to mitigate thermocapillary effects and facilitate optical alignment for automated image capture and signal analysis. The system-on-board prototype, coupled with the LAMP primers designed by BioCoS, was sensitive enough to detect large variations in viral loads of SARS-CoV-2 corresponding to a threshold cycle range of 16 to 39. Furthermore, tested samples consisted of a broad range of viral strains and lineages identified in Canada during 2021–2022. Clinical specimens were collected and tested at the Kingston Health Science Centre using a clinically validated PCR assay, and variants were determined using whole genome sequencing.
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spelling pubmed-103776822023-07-29 A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples Staples, Jake Dourou, Athanasia-Maria Liampa, Irene Sjaarda, Calvin Moslinger, Emily Wong, Henry Sheth, Prameet M. Arhondakis, Stilianos Prakash, Ravi Biomedicines Article We report a small-footprint cost-effective isothermal rapid DNA amplification system, with integrated microfluidics for automated sample analysis and detection of SARS-CoV-2 in human and environmental samples. Our system measures low-level fluorescent signals in real-time during amplification, while maintaining the desired assay temperature on a low power, portable system footprint. A unique soft microfluidic chip design was implemented to mitigate thermocapillary effects and facilitate optical alignment for automated image capture and signal analysis. The system-on-board prototype, coupled with the LAMP primers designed by BioCoS, was sensitive enough to detect large variations in viral loads of SARS-CoV-2 corresponding to a threshold cycle range of 16 to 39. Furthermore, tested samples consisted of a broad range of viral strains and lineages identified in Canada during 2021–2022. Clinical specimens were collected and tested at the Kingston Health Science Centre using a clinically validated PCR assay, and variants were determined using whole genome sequencing. MDPI 2023-07-20 /pmc/articles/PMC10377682/ /pubmed/37509680 http://dx.doi.org/10.3390/biomedicines11072038 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
Staples, Jake
Dourou, Athanasia-Maria
Liampa, Irene
Sjaarda, Calvin
Moslinger, Emily
Wong, Henry
Sheth, Prameet M.
Arhondakis, Stilianos
Prakash, Ravi
A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title_full A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title_fullStr A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title_full_unstemmed A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title_short A Miniaturized System for Rapid, Isothermal Detection of SARS-CoV-2 in Human and Environmental Samples
title_sort miniaturized system for rapid, isothermal detection of sars-cov-2 in human and environmental samples
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10377682/
https://www.ncbi.nlm.nih.gov/pubmed/37509680
http://dx.doi.org/10.3390/biomedicines11072038
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