Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples
Open Access
- 19 January 2021
- journal article
- research article
- Published by Springer Science and Business Media LLC in Scientific Reports
- Vol. 11 (1), 1-10
- https://doi.org/10.1038/s41598-020-80352-8
Abstract
RT-LAMP detection of SARS-CoV-2 has been shown to be a valuable approach to scale up COVID-19 diagnostics and thus contribute to limiting the spread of the disease. Here we present the optimization of highly cost-effective in-house produced enzymes, and we benchmark their performance against commercial alternatives. We explore the compatibility between multiple DNA polymerases with high strand-displacement activity and thermostable reverse transcriptases required for RT-LAMP. We optimize reaction conditions and demonstrate their applicability using both synthetic RNA and clinical patient samples. Finally, we validate the optimized RT-LAMP assay for the detection of SARS-CoV-2 in unextracted heat-inactivated nasopharyngeal samples from 184 patients. We anticipate that optimized and affordable reagents for RT-LAMP will facilitate the expansion of SARS-CoV-2 testing globally, especially in sites and settings where the need for large scale testing cannot be met by commercial alternatives.Funding Information
- Natural Science Foundation of Liaoning Province (1580441949000)
- Ministry of Science and Technology of the People’s Republic of China (SQ2020YFF0411358)
- Ganzhou COVID-19 Emergency Research Project
- Knut och Alice Wallenbergs Stiftelse (KAW 2020.0182, 2016.0123)
- Ragnar Söderbergs stiftelse (Swedish Foundations' Starting Grant)
- Vetenskapsrådet (VR 2016-01842)
- Swedish Foundation for International Cooperation in Research and Higher Education (CH2018-7750)
- Karolinska Institutet (SciLifeLab Fellowship, SFO, KID and KI funds)
- Karolinska Institute
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