Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples.
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Alekseenko A, Barrett D, Pareja-Sanchez Y, Howard RJ, Strandback E, Ampah-Korsah H, Rovšnik U, Zuniga-Veliz S, Klenov A, Malloo J, Ye S, Liu X, Reinius B, Elsässer SJ, Nyman T, Sandh G, Yin X, Pelechano V
Sci Rep 11 (1) 1820 [2021-01-19; online 2021-01-19]
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.
PubMed 33469065
DOI 10.1038/s41598-020-80352-8
Crossref 10.1038/s41598-020-80352-8
pii: 10.1038/s41598-020-80352-8
pmc: PMC7815738