Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples
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 agains...
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Nature Portfolio
2021
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oai:doaj.org-article:1f27a366d4e94aed943f786f5264f7a02021-12-02T13:51:16ZDirect detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples10.1038/s41598-020-80352-82045-2322https://doaj.org/article/1f27a366d4e94aed943f786f5264f7a02021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-80352-8https://doaj.org/toc/2045-2322Abstract 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.Alisa AlekseenkoDonal BarrettYerma Pareja-SanchezRebecca J. HowardEmilia StrandbackHenry Ampah-KorsahUrška RovšnikSilvia Zuniga-VelizAlexander KlenovJayshna MallooShenglong YeXiyang LiuBjörn ReiniusSimon J. ElsässerTomas NymanGustaf SandhXiushan YinVicent PelechanoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-10 (2021) |
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Medicine R Science Q Alisa Alekseenko Donal Barrett Yerma Pareja-Sanchez Rebecca J. Howard Emilia Strandback Henry Ampah-Korsah Urška Rovšnik Silvia Zuniga-Veliz Alexander Klenov Jayshna Malloo Shenglong Ye Xiyang Liu Björn Reinius Simon J. Elsässer Tomas Nyman Gustaf Sandh Xiushan Yin Vicent Pelechano Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| description |
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. |
| format |
article |
| author |
Alisa Alekseenko Donal Barrett Yerma Pareja-Sanchez Rebecca J. Howard Emilia Strandback Henry Ampah-Korsah Urška Rovšnik Silvia Zuniga-Veliz Alexander Klenov Jayshna Malloo Shenglong Ye Xiyang Liu Björn Reinius Simon J. Elsässer Tomas Nyman Gustaf Sandh Xiushan Yin Vicent Pelechano |
| author_facet |
Alisa Alekseenko Donal Barrett Yerma Pareja-Sanchez Rebecca J. Howard Emilia Strandback Henry Ampah-Korsah Urška Rovšnik Silvia Zuniga-Veliz Alexander Klenov Jayshna Malloo Shenglong Ye Xiyang Liu Björn Reinius Simon J. Elsässer Tomas Nyman Gustaf Sandh Xiushan Yin Vicent Pelechano |
| author_sort |
Alisa Alekseenko |
| title |
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| title_short |
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| title_full |
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| title_fullStr |
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| title_full_unstemmed |
Direct detection of SARS-CoV-2 using non-commercial RT-LAMP reagents on heat-inactivated samples |
| title_sort |
direct detection of sars-cov-2 using non-commercial rt-lamp reagents on heat-inactivated samples |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/1f27a366d4e94aed943f786f5264f7a0 |
| work_keys_str_mv |
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