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Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples
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Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples
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Journal Article
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples
2021
Overview
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.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ COVID-19
/ Humanities and Social Sciences
/ Humans
/ Molecular Diagnostic Techniques - methods
/ Nucleic Acid Amplification Techniques - methods
/ Reagents
/ RNA
/ RNA-Directed DNA Polymerase - metabolism
/ SARS-CoV-2 - isolation & purification
/ Science
