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An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system
Outbreaks of both influenza virus and the novel coronavirus SARS-CoV-2 are serious threats to human health and life. It is very important to establish a rapid, accurate test with large-scale detection potential to prevent the further spread of the epidemic. An optimized RPA-Cas12a-based platform com...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Science China Press
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8809245/ https://www.ncbi.nlm.nih.gov/pubmed/35126481 http://dx.doi.org/10.1007/s11426-021-1169-1 |
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author | Sun, Zhen Lin, Kang-Feng Zhao, Ze-Hang Wang, Yang Hong, Xin-Xin Guo, Jian-Guang Ruan, Qing-Yu Lu, Lian-Yu Li, Xiao Zhang, Rui Yang, Chao-Yong Li, Bo-An |
author_facet | Sun, Zhen Lin, Kang-Feng Zhao, Ze-Hang Wang, Yang Hong, Xin-Xin Guo, Jian-Guang Ruan, Qing-Yu Lu, Lian-Yu Li, Xiao Zhang, Rui Yang, Chao-Yong Li, Bo-An |
author_sort | Sun, Zhen |
collection | PubMed |
description | Outbreaks of both influenza virus and the novel coronavirus SARS-CoV-2 are serious threats to human health and life. It is very important to establish a rapid, accurate test with large-scale detection potential to prevent the further spread of the epidemic. An optimized RPA-Cas12a-based platform combined with digital microfluidics (DMF), the RCD platform, was established to achieve the automated, rapid detection of influenza viruses and SARS-CoV-2. The probe in the RPA-Cas12a system was optimized to produce maximal fluorescence to increase the amplification signal. The reaction droplets in the platform were all at the microliter level and the detection could be accomplished within 30 min due to the effective mixing of droplets by digital microfluidic technology. The whole process from amplification to recognition is completed in the chip, which reduces the risk of aerosol contamination. One chip can contain multiple detection reaction areas, offering the potential for customized detection. The RCD platform demonstrated a high level of sensitivity, specificity (no false positives or negatives), speed (≤30 min), automation and multiplexing. We also used the RCD platform to detect nucleic acids from influenza patients and COVID-19 patients. The results were consistent with the findings of qPCR. The RCD platform is a one-step, rapid, highly sensitive and specific method with the advantages of digital microfluidic technology, which circumvents the shortcomings of manual operation. The development of the RCD platform provides potential for the isothermal automatic detection of nucleic acids during epidemics. [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available in the online version of this article at 10.1007/s11426-021-1169-1. |
format | Online Article Text |
id | pubmed-8809245 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Science China Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-88092452022-02-02 An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system Sun, Zhen Lin, Kang-Feng Zhao, Ze-Hang Wang, Yang Hong, Xin-Xin Guo, Jian-Guang Ruan, Qing-Yu Lu, Lian-Yu Li, Xiao Zhang, Rui Yang, Chao-Yong Li, Bo-An Sci China Chem Articles Outbreaks of both influenza virus and the novel coronavirus SARS-CoV-2 are serious threats to human health and life. It is very important to establish a rapid, accurate test with large-scale detection potential to prevent the further spread of the epidemic. An optimized RPA-Cas12a-based platform combined with digital microfluidics (DMF), the RCD platform, was established to achieve the automated, rapid detection of influenza viruses and SARS-CoV-2. The probe in the RPA-Cas12a system was optimized to produce maximal fluorescence to increase the amplification signal. The reaction droplets in the platform were all at the microliter level and the detection could be accomplished within 30 min due to the effective mixing of droplets by digital microfluidic technology. The whole process from amplification to recognition is completed in the chip, which reduces the risk of aerosol contamination. One chip can contain multiple detection reaction areas, offering the potential for customized detection. The RCD platform demonstrated a high level of sensitivity, specificity (no false positives or negatives), speed (≤30 min), automation and multiplexing. We also used the RCD platform to detect nucleic acids from influenza patients and COVID-19 patients. The results were consistent with the findings of qPCR. The RCD platform is a one-step, rapid, highly sensitive and specific method with the advantages of digital microfluidic technology, which circumvents the shortcomings of manual operation. The development of the RCD platform provides potential for the isothermal automatic detection of nucleic acids during epidemics. [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: Supplementary material is available in the online version of this article at 10.1007/s11426-021-1169-1. Science China Press 2022-01-25 2022 /pmc/articles/PMC8809245/ /pubmed/35126481 http://dx.doi.org/10.1007/s11426-021-1169-1 Text en © Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2022 This article is made available via the PMC Open Access Subset for unrestricted research re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Articles Sun, Zhen Lin, Kang-Feng Zhao, Ze-Hang Wang, Yang Hong, Xin-Xin Guo, Jian-Guang Ruan, Qing-Yu Lu, Lian-Yu Li, Xiao Zhang, Rui Yang, Chao-Yong Li, Bo-An An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title | An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title_full | An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title_fullStr | An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title_full_unstemmed | An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title_short | An automated nucleic acid detection platform using digital microfluidics with an optimized Cas12a system |
title_sort | automated nucleic acid detection platform using digital microfluidics with an optimized cas12a system |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8809245/ https://www.ncbi.nlm.nih.gov/pubmed/35126481 http://dx.doi.org/10.1007/s11426-021-1169-1 |
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