CoLAMP : CRISPR-based one-pot loop-mediated isothermal amplification enables at-home diagnosis of SARS-CoV-2 RNA with nearly eliminated contamination utilizing amplicons depletion strategy
Copyright © 2023 Elsevier B.V. All rights reserved..
Rapid point-of-care diagnostics, essential in settings such as airport on-site testing and home-based screening, displayed important implications for infectious disease control during the SARS-CoV-2 outbreak. However, the deployment of simple and sensitive assays in real-life scenarios still faces the concern of aerosol contamination. Here, we report an amplicon-depleting CRISPR-based one-pot loop-mediated isothermal amplification (CoLAMP) assay for point-of-care diagnosis of SARS-CoV-2 RNA. In this work, AapCas12b sgRNA is designed to recognize the activator sequence sited in the loop region of the LAMP product, which is crucial for exponential amplification. By destroying the aerosol-prone amplifiable products at the end of each amplification reaction, our design can significantly reduce the amplicons contamination that causes false positive results in point-of-care diagnostics. For at-home self-testing, we designed a low-cost sample-to-result device for fluorescence-based visual interpretation. As well, a commercial portable electrochemical platform was deployed as a proof-of-concept of ready-to-use point-of-care diagnostic systems. The field deployable CoLAMP assay can detect as low as 0.5 copies/μL of SARS-CoV-2 RNA in clinical nasopharyngeal swab samples within 40 min without the need for specialists for its operation.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:236 |
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| Enthalten in: |
Biosensors & bioelectronics - 236(2023) vom: 15. Sept., Seite 115402 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Cao, Yumeng [VerfasserIn] |
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| Links: |
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| Themen: |
CRISPR-Based diagnostics |
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| Anmerkungen: |
Date Completed 19.06.2023 Date Revised 19.06.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.bios.2023.115402 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM357463048 |
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| 500 | |a published: Print-Electronic | ||
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| 520 | |a Copyright © 2023 Elsevier B.V. All rights reserved. | ||
| 520 | |a Rapid point-of-care diagnostics, essential in settings such as airport on-site testing and home-based screening, displayed important implications for infectious disease control during the SARS-CoV-2 outbreak. However, the deployment of simple and sensitive assays in real-life scenarios still faces the concern of aerosol contamination. Here, we report an amplicon-depleting CRISPR-based one-pot loop-mediated isothermal amplification (CoLAMP) assay for point-of-care diagnosis of SARS-CoV-2 RNA. In this work, AapCas12b sgRNA is designed to recognize the activator sequence sited in the loop region of the LAMP product, which is crucial for exponential amplification. By destroying the aerosol-prone amplifiable products at the end of each amplification reaction, our design can significantly reduce the amplicons contamination that causes false positive results in point-of-care diagnostics. For at-home self-testing, we designed a low-cost sample-to-result device for fluorescence-based visual interpretation. As well, a commercial portable electrochemical platform was deployed as a proof-of-concept of ready-to-use point-of-care diagnostic systems. The field deployable CoLAMP assay can detect as low as 0.5 copies/μL of SARS-CoV-2 RNA in clinical nasopharyngeal swab samples within 40 min without the need for specialists for its operation | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a CRISPR-Based diagnostics | |
| 650 | 4 | |a Contamination eliminated | |
| 650 | 4 | |a LAMP | |
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| 700 | 1 | |a Lin, Haosi |e verfasserin |4 aut | |
| 700 | 1 | |a Rodriguez Serrano, Alan Fernando |e verfasserin |4 aut | |
| 700 | 1 | |a Lui, Grace C Y |e verfasserin |4 aut | |
| 700 | 1 | |a Hsing, I-Ming |e verfasserin |4 aut | |
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