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.
【저자키워드】 viral infection, PCR-based techniques, 【초록키워드】 COVID-19, SARS-CoV-2, diagnostics, RNA, Spread, RT-LAMP, SARS-CoV-2 testing, Patient, expansion, Enzymes, optimization, patients, reverse transcriptase, reverse transcriptases, house, Nasopharyngeal samples, applicability, compatibility, reagent, DNA polymerase, nasopharyngeal sample, heat-inactivated, approach, produced, shown, the disease, required, facilitate, condition, contribute, 【제목키워드】 SARS-CoV-2, reagent, Direct, heat-inactivated,