Abstract
AbstractRT-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.
Funder
Natural Science Foundation of Liaoning Province
Ministry of Science and Technology of the People’s Republic of China
Ganzhou COVID-19 Emergency Research Project
Knut och Alice Wallenbergs Stiftelse
Ragnar Söderbergs stiftelse
Vetenskapsrådet
Swedish Foundation for International Cooperation in Research and Higher Education
Karolinska Institutet
Karolinska Institute
Publisher
Springer Science and Business Media LLC
Cited by
38 articles.
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