Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP
Abstract Rapid, reliable, and widespread testing is required to curtail the ongoing COVID-19 pandemic. Current gold standard nucleic acid tests are hampered by supply shortages in critical reagents including nasal swabs, RNA extraction kits, personal protective equipment (PPE), instrumentation, and labor. Here we present an approach to overcome these challenges with the development of a rapid colorimetric assay using reverse-transcription loop-mediated isothermal amplification (RT-LAMP) optimized on human saliva samples without an RNA purification step. We describe our optimizations of the LAMP reaction and saliva pretreatment protocols that enabled rapid and sensitive detection of < 102 viral genomes per reaction in contrived saliva controls. Moreover, our saliva pretreatment protocol enabled sensitive viral detection by conventional quantitative reverse transcription polymerase chain reaction (qRT-PCR) without RNA extraction. We validated the high performance of these assays on clinical samples and demonstrate a promising approach to overcome the current bottlenecks limiting widespread testing..
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Preprint| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
bioRxiv.org - (2021) vom: 23. Jan. Zur Gesamtaufnahme - year:2021 |
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| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lalli, Matthew A. [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| doi: |
10.1101/2020.05.07.20093542 |
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| funding: |
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| PPN (Katalog-ID): |
XBI01777666X |
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| 245 | 1 | 0 | |a Rapid and extraction-free detection of SARS-CoV-2 from saliva with colorimetric LAMP |
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| 520 | |a Abstract Rapid, reliable, and widespread testing is required to curtail the ongoing COVID-19 pandemic. Current gold standard nucleic acid tests are hampered by supply shortages in critical reagents including nasal swabs, RNA extraction kits, personal protective equipment (PPE), instrumentation, and labor. Here we present an approach to overcome these challenges with the development of a rapid colorimetric assay using reverse-transcription loop-mediated isothermal amplification (RT-LAMP) optimized on human saliva samples without an RNA purification step. We describe our optimizations of the LAMP reaction and saliva pretreatment protocols that enabled rapid and sensitive detection of < 102 viral genomes per reaction in contrived saliva controls. Moreover, our saliva pretreatment protocol enabled sensitive viral detection by conventional quantitative reverse transcription polymerase chain reaction (qRT-PCR) without RNA extraction. We validated the high performance of these assays on clinical samples and demonstrate a promising approach to overcome the current bottlenecks limiting widespread testing. | ||
| 700 | 1 | |a Langmade, S. Joshua |e verfasserin |4 aut | |
| 700 | 1 | |a Chen, Xuhua |e verfasserin |4 aut | |
| 700 | 1 | |a Fronick, Catrina C. |e verfasserin |4 aut | |
| 700 | 1 | |a Sawyer, Christopher S. |e verfasserin |4 aut | |
| 700 | 1 | |a Burcea, Lauren C. |e verfasserin |4 aut | |
| 700 | 1 | |a Wilkinson, Michael N. |e verfasserin |4 aut | |
| 700 | 1 | |a Fulton, Robert S. |e verfasserin |4 aut | |
| 700 | 1 | |a Heinz, Michael |e verfasserin |4 aut | |
| 700 | 1 | |a Buchser, William J. |e verfasserin |4 aut | |
| 700 | 1 | |a Head, Richard D. |e verfasserin |4 aut | |
| 700 | 1 | |a Mitra, Robi D. |e verfasserin |4 aut | |
| 700 | 1 | |a Milbrandt, Jeffrey |e verfasserin |4 aut | |
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