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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2022
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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. |
format | Online Article Text |
id | pubmed-8653405 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
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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