Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2
Summary: The COVID-19 pandemic has spread worldwide, and rapid detection of the SARS-CoV-2 virus is crucial for infection surveillance and epidemic control. This study developed a centrifugal microfluidics-based multiplex reverse transcription recombinase polymerase amplification (RT-RPA) assay for...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-03-01
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| Series: | iScience |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S258900422300322X |
| _version_ | 1811159919197421568 |
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| author | Ruoxu Li Ning Su Xiaodong Ren Xiange Sun Wenman Li Yuwei Li Jin Li Chen Chen Hong Wang Weiping Lu Shaoli Deng Qing Huang |
| author_facet | Ruoxu Li Ning Su Xiaodong Ren Xiange Sun Wenman Li Yuwei Li Jin Li Chen Chen Hong Wang Weiping Lu Shaoli Deng Qing Huang |
| author_sort | Ruoxu Li |
| collection | DOAJ |
| description | Summary: The COVID-19 pandemic has spread worldwide, and rapid detection of the SARS-CoV-2 virus is crucial for infection surveillance and epidemic control. This study developed a centrifugal microfluidics-based multiplex reverse transcription recombinase polymerase amplification (RT-RPA) assay for endpoint fluorescence detection of the E, N, and ORF1ab genes of SARS-CoV-2. The microscope slide-shaped microfluidic chip could simultaneously accomplish three target genes and one reference human gene (i.e., ACTB) RT-RPA reactions in 30 min, and the sensitivity was 40 RNA copies/reaction for the E gene, 20 RNA copies/reaction for the N gene, and 10 RNA copies/reaction for the ORF1ab gene. The chip demonstrated high specificity, reproducibility, and repeatability. Chip performance was also evaluated using real clinical samples. Thus, this rapid, accurate, on-site, and multiplexed nucleic acid test microfluidic chip would significantly contribute to detecting patients with COVID-19 in low-resource settings and point-of-care testing (POCT) and, in the future, could be used to detect emerging new variants of SARS-CoV-2. |
| first_indexed | 2024-04-10T05:49:55Z |
| format | Article |
| id | doaj.art-a165215664514943ad86e2f1790535ad |
| institution | Directory Open Access Journal |
| issn | 2589-0042 |
| language | English |
| last_indexed | 2024-04-10T05:49:55Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj.art-a165215664514943ad86e2f1790535ad2023-03-05T04:25:13ZengElsevieriScience2589-00422023-03-01263106245Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2Ruoxu Li0Ning Su1Xiaodong Ren2Xiange Sun3Wenman Li4Yuwei Li5Jin Li6Chen Chen7Hong Wang8Weiping Lu9Shaoli Deng10Qing Huang11Department of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, ChinaDepartment of Laboratory Medicine, Daping Hospital, Army Medical University, Chongqing 400042, China; Corresponding authorSummary: The COVID-19 pandemic has spread worldwide, and rapid detection of the SARS-CoV-2 virus is crucial for infection surveillance and epidemic control. This study developed a centrifugal microfluidics-based multiplex reverse transcription recombinase polymerase amplification (RT-RPA) assay for endpoint fluorescence detection of the E, N, and ORF1ab genes of SARS-CoV-2. The microscope slide-shaped microfluidic chip could simultaneously accomplish three target genes and one reference human gene (i.e., ACTB) RT-RPA reactions in 30 min, and the sensitivity was 40 RNA copies/reaction for the E gene, 20 RNA copies/reaction for the N gene, and 10 RNA copies/reaction for the ORF1ab gene. The chip demonstrated high specificity, reproducibility, and repeatability. Chip performance was also evaluated using real clinical samples. Thus, this rapid, accurate, on-site, and multiplexed nucleic acid test microfluidic chip would significantly contribute to detecting patients with COVID-19 in low-resource settings and point-of-care testing (POCT) and, in the future, could be used to detect emerging new variants of SARS-CoV-2.http://www.sciencedirect.com/science/article/pii/S258900422300322XBiological sciencesBiotechnologyFluidicsNucleic acids |
| spellingShingle | Ruoxu Li Ning Su Xiaodong Ren Xiange Sun Wenman Li Yuwei Li Jin Li Chen Chen Hong Wang Weiping Lu Shaoli Deng Qing Huang Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 iScience Biological sciences Biotechnology Fluidics Nucleic acids |
| title | Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 |
| title_full | Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 |
| title_fullStr | Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 |
| title_full_unstemmed | Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 |
| title_short | Centrifugal microfluidic-based multiplex recombinase polymerase amplification assay for rapid detection of SARS-CoV-2 |
| title_sort | centrifugal microfluidic based multiplex recombinase polymerase amplification assay for rapid detection of sars cov 2 |
| topic | Biological sciences Biotechnology Fluidics Nucleic acids |
| url | http://www.sciencedirect.com/science/article/pii/S258900422300322X |
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