The DOI matches its Crossref record; 34 of the 34 references it cites were found in a registry or an index.
Rapid and Extraction-Free Detection of SARS-CoV-2 from Saliva by Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification
Open accessAbstract
Background: 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, instrumentation, and labor. Methods: To overcome these challenges, we developed 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 the optimization of saliva pretreatment protocols to enable analytically sensitive viral detection by RT-LAMP. We optimized the RT-LAMP reaction conditions and implemented high-throughput unbiased methods for assay interpretation. We tested whether saliva pretreatment could also enable viral detection by conventional reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Finally, we validated these assays on clinical samples. Results: The optimized saliva pretreatment protocol enabled analytically sensitive extraction-free detection of SARS-CoV-2 from saliva by colorimetric RT-LAMP or RT-qPCR. In simulated samples, the optimized RT-LAMP assay had a limit of detection of 59 (95% confidence interval: 44–104) particle copies per reaction. We highlighted the flexibility of LAMP assay implementation using 3 readouts: naked-eye colorimetry, spectrophotometry, and real-time fluorescence. In a set of 30 clinical saliva samples, colorimetric RT-LAMP and RT-qPCR assays performed directly on pretreated saliva samples without RNA extraction had accuracies greater than 90%. Conclusions: Rapid and extraction-free detection of SARS-CoV-2 from saliva by colorimetric RT-LAMP is a simple, sensitive, and cost-effective approach with broad potential to expand diagnostic testing for the virus causing COVID-19.
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Cites
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Test sensitivity is secondary to frequency and turnaround time for COVID-19 screening
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Saliva or Nasopharyngeal Swab Specimens for Detection of SARS-CoV-2
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Cited by
Rapid, point-of-care antigen and molecular-based tests for diagnosis of SARS-CoV-2 infection
Dinnes, J., Deeks, J. J., Adriano, A., Berhane, S., Davenport, C., Dittrich, S., Emperador, D., Takwoingi, Y., Cunningham, J., Beese, S., et al. (2020) · Cochrane Database of Systematic Reviews · doi:10.1002/14651858.cd013705 · Cited by 1,156
Considerations for diagnostic COVID-19 tests
Vandenberg, O., Martiny, D., Rochas, O., van Belkum, A., & Kozlakidis, Z. (2021) · Nature Reviews Microbiology · doi:10.1038/s41579-020-00461-z · Cited by 803
Rapid, point-of-care antigen tests for diagnosis of SARS-CoV-2 infection
Dinnes, J., Deeks, J. J., Berhane, S., Taylor, M., Adriano, A., Davenport, C., Dittrich, S., Emperador, D., Takwoingi, Y., Cunningham, J., et al. (2021) · Cochrane Database of Systematic Reviews · doi:10.1002/14651858.cd013705.pub2 · Cited by 591
Lamb, L. E., Bartolone, S. N., Ward, E., & Chancellor, M. B. (2020) · PLOS ONE · doi:10.1371/journal.pone.0234682 · Cited by 348
Najjar, D., Rainbow, J., Sharma Timilsina, S., Jolly, P., de Puig, H., Yafia, M., Durr, N., Sallum, H., Alter, G., Li, J. Z., et al. (2022) · Nature Biomedical Engineering · doi:10.1038/s41551-022-00919-w · Cited by 291
Metadata
- Authors
- Lalli, M. A., Langmade, J. S., Chen, X., Fronick, C. C., Sawyer, C. S., Burcea, L. C., Wilkinson, M. N., Fulton, R. S., Heinz, M., Buchser, W. J., Head, R. D., Mitra, R. D., & Milbrandt, J.
- Venue
- Clinical Chemistry
- Publisher
- Oxford University Press (OUP)
- Published
- 2020-10-24
- Volume
- 67
- Issue
- 2
- Pages
- 415-424
- ISSN
- 0009-9147 (ISSN-L)
- 1530-8561
- OpenAlex
- W3093690976
- Cited by
- 267
- References
- 34
- FWCI
- 6.61
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Lalli, M. A., Langmade, J. S., Chen, X., Fronick, C. C., Sawyer, C. S., Burcea, L. C., Wilkinson, M. N., Fulton, R. S., Heinz, M., Buchser, W. J., Head, R. D., Mitra, R. D., & Milbrandt, J. (2021). Rapid and Extraction-Free Detection of SARS-CoV-2 from Saliva by Colorimetric Reverse-Transcription Loop-Mediated Isothermal Amplification. Clinical Chemistry. https://doi.org/10.1093/clinchem/hvaa267
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
