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Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system

Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we deve...

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Autores principales: Li, Ziyue, Ding, Xiong, Yin, Kun, Avery, Lori, Ballesteros, Enrique, Liu, Changchun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8653405/
https://www.ncbi.nlm.nih.gov/pubmed/34906838
http://dx.doi.org/10.1016/j.bios.2021.113865
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author Li, Ziyue
Ding, Xiong
Yin, Kun
Avery, Lori
Ballesteros, Enrique
Liu, Changchun
author_facet Li, Ziyue
Ding, Xiong
Yin, Kun
Avery, Lori
Ballesteros, Enrique
Liu, Changchun
author_sort Li, Ziyue
collection PubMed
description Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we developed a simple, sensitive, instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using a self-contained microfluidic system. The microfluidic chip integrates isothermal amplification, CRISPR cleavage, and lateral flow detection in a single, closed microfluidic platform, enabling contamination-free, visual detection. To simplify the operation and transportation of the device, we lyophilized the CRISPR reagents in the reaction chamber and pre-stored the liquid solutions in blisters. We employed a low-cost, portable hand warmer to incubate the microfluidic chip without the need for electricity. The self-contained microfluidic system can detect down to 100 copies of SARS-CoV-2 RNA. Further, we clinically validated our method by detecting 24 COVID-19 clinical nasopharyngeal swab samples, achieving excellent sensitivity (94.1%), specificity (100%), and accuracy (95.8%). This simple, sensitive, and affordable microfluidic system represents a promising tool for point-of-care diagnostics of COVID-19 and other infectious diseases.
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spelling pubmed-86534052021-12-08 Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system Li, Ziyue Ding, Xiong Yin, Kun Avery, Lori Ballesteros, Enrique Liu, Changchun Biosens Bioelectron Article Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we developed a simple, sensitive, instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using a self-contained microfluidic system. The microfluidic chip integrates isothermal amplification, CRISPR cleavage, and lateral flow detection in a single, closed microfluidic platform, enabling contamination-free, visual detection. To simplify the operation and transportation of the device, we lyophilized the CRISPR reagents in the reaction chamber and pre-stored the liquid solutions in blisters. We employed a low-cost, portable hand warmer to incubate the microfluidic chip without the need for electricity. The self-contained microfluidic system can detect down to 100 copies of SARS-CoV-2 RNA. Further, we clinically validated our method by detecting 24 COVID-19 clinical nasopharyngeal swab samples, achieving excellent sensitivity (94.1%), specificity (100%), and accuracy (95.8%). This simple, sensitive, and affordable microfluidic system represents a promising tool for point-of-care diagnostics of COVID-19 and other infectious diseases. Elsevier B.V. 2022-03-01 2021-12-08 /pmc/articles/PMC8653405/ /pubmed/34906838 http://dx.doi.org/10.1016/j.bios.2021.113865 Text en © 2021 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Article
Li, Ziyue
Ding, Xiong
Yin, Kun
Avery, Lori
Ballesteros, Enrique
Liu, Changchun
Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title_full Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title_fullStr Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title_full_unstemmed Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title_short Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system
title_sort instrument-free, crispr-based diagnostics of sars-cov-2 using self-contained microfluidic system
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8653405/
https://www.ncbi.nlm.nih.gov/pubmed/34906838
http://dx.doi.org/10.1016/j.bios.2021.113865
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