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Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination
Continuous mutations have occurred in the genome of the SARS-CoV-2 virus since the onset of the COVID-19 pandemic. The increased transmissibility of the mutated viruses has not only imposed medical burdens but also prolonged the duration of the pandemic. A point-of-care (POC) platform that provides...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10456927/ https://www.ncbi.nlm.nih.gov/pubmed/37630161 http://dx.doi.org/10.3390/mi14081627 |
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author | Ho, Kuan-Lun Ding, Jing Fan, Jia-Shao Tsui, Wai Ning Tiffany Bai, Jianfa Fan, Shih-Kang |
author_facet | Ho, Kuan-Lun Ding, Jing Fan, Jia-Shao Tsui, Wai Ning Tiffany Bai, Jianfa Fan, Shih-Kang |
author_sort | Ho, Kuan-Lun |
collection | PubMed |
description | Continuous mutations have occurred in the genome of the SARS-CoV-2 virus since the onset of the COVID-19 pandemic. The increased transmissibility of the mutated viruses has not only imposed medical burdens but also prolonged the duration of the pandemic. A point-of-care (POC) platform that provides multitarget detection will help to track and reduce disease transmissions. Here we detected and discriminated three genotypes of SARS-CoV-2, including the wildtype and two variants of concern (VOCs), the Delta variant and Omicron variant, through reverse transcription quantitative polymerase chain reaction (RT-qPCR) on a digital microfluidics (DMF)-based cartridge. Upon evaluating with the RNA samples of Omicron variant, the DMF RT-qPCR presented a sensitivity of 10 copies/μL and an amplification efficiency of 96.1%, capable for clinical diagnosis. When spiking with SARS-CoV-2 RNA (wildtype, Delta variant, or Omicron variant) and 18S rDNA, the clinical analog samples demonstrated accurate detection and discrimination of different SARS-CoV-2 strains in 49 min. |
format | Online Article Text |
id | pubmed-10456927 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-104569272023-08-26 Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination Ho, Kuan-Lun Ding, Jing Fan, Jia-Shao Tsui, Wai Ning Tiffany Bai, Jianfa Fan, Shih-Kang Micromachines (Basel) Article Continuous mutations have occurred in the genome of the SARS-CoV-2 virus since the onset of the COVID-19 pandemic. The increased transmissibility of the mutated viruses has not only imposed medical burdens but also prolonged the duration of the pandemic. A point-of-care (POC) platform that provides multitarget detection will help to track and reduce disease transmissions. Here we detected and discriminated three genotypes of SARS-CoV-2, including the wildtype and two variants of concern (VOCs), the Delta variant and Omicron variant, through reverse transcription quantitative polymerase chain reaction (RT-qPCR) on a digital microfluidics (DMF)-based cartridge. Upon evaluating with the RNA samples of Omicron variant, the DMF RT-qPCR presented a sensitivity of 10 copies/μL and an amplification efficiency of 96.1%, capable for clinical diagnosis. When spiking with SARS-CoV-2 RNA (wildtype, Delta variant, or Omicron variant) and 18S rDNA, the clinical analog samples demonstrated accurate detection and discrimination of different SARS-CoV-2 strains in 49 min. MDPI 2023-08-17 /pmc/articles/PMC10456927/ /pubmed/37630161 http://dx.doi.org/10.3390/mi14081627 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 Ho, Kuan-Lun Ding, Jing Fan, Jia-Shao Tsui, Wai Ning Tiffany Bai, Jianfa Fan, Shih-Kang Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title | Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title_full | Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title_fullStr | Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title_full_unstemmed | Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title_short | Digital Microfluidic Multiplex RT-qPCR for SARS-CoV-2 Detection and Variants Discrimination |
title_sort | digital microfluidic multiplex rt-qpcr for sars-cov-2 detection and variants discrimination |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10456927/ https://www.ncbi.nlm.nih.gov/pubmed/37630161 http://dx.doi.org/10.3390/mi14081627 |
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