Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction
Abstract The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) t...
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Nature Portfolio
2021
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oai:doaj.org-article:6d182b455806472a8759a067b2e4669f2021-12-02T13:57:37ZDirect diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction10.1038/s41598-021-81487-y2045-2322https://doaj.org/article/6d182b455806472a8759a067b2e4669f2021-01-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-81487-yhttps://doaj.org/toc/2045-2322Abstract The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) that uses readily available reagents and a simple heat block using contrived spike-in and actual clinical samples. RT-LAMP testing on RNA-spiked samples showed a limit of detection (LoD) of 2.5 copies/μl of viral transport media. RT-LAMP testing directly on clinical nasopharyngeal swab samples in viral transport media had an 85% positive percentage agreement (PPA) (17/20), and 100% negative percentage agreement (NPV) and delivered results in 30 min. Our optimized RT-LAMP based testing method is a scalable system that is sufficiently sensitive and robust to test for SARS-CoV-2 directly on clinical nasopharyngeal swab samples in viral transport media in 30 min at the point of care without the need for specialized or proprietary equipment or reagents. This cost-effective and efficient one-step testing method can be readily available for COVID-19 testing world-wide, especially in resource poor settings.Shan WeiEsther KohlAlexandre DjandjiStephanie MorganSusan WhittierMahesh MansukhaniEldad HodMary D’AltonYousin SuhZev WilliamsNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-6 (2021) |
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DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
Medicine R Science Q |
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Medicine R Science Q Shan Wei Esther Kohl Alexandre Djandji Stephanie Morgan Susan Whittier Mahesh Mansukhani Eldad Hod Mary D’Alton Yousin Suh Zev Williams Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| description |
Abstract The COVID-19 pandemic has resulted in an urgent need for a rapid, point of care diagnostic testing that could be rapidly scaled on a worldwide level. We developed and tested a highly sensitive and robust assay based on reverse transcription loop mediated isothermal amplification (RT-LAMP) that uses readily available reagents and a simple heat block using contrived spike-in and actual clinical samples. RT-LAMP testing on RNA-spiked samples showed a limit of detection (LoD) of 2.5 copies/μl of viral transport media. RT-LAMP testing directly on clinical nasopharyngeal swab samples in viral transport media had an 85% positive percentage agreement (PPA) (17/20), and 100% negative percentage agreement (NPV) and delivered results in 30 min. Our optimized RT-LAMP based testing method is a scalable system that is sufficiently sensitive and robust to test for SARS-CoV-2 directly on clinical nasopharyngeal swab samples in viral transport media in 30 min at the point of care without the need for specialized or proprietary equipment or reagents. This cost-effective and efficient one-step testing method can be readily available for COVID-19 testing world-wide, especially in resource poor settings. |
| format |
article |
| author |
Shan Wei Esther Kohl Alexandre Djandji Stephanie Morgan Susan Whittier Mahesh Mansukhani Eldad Hod Mary D’Alton Yousin Suh Zev Williams |
| author_facet |
Shan Wei Esther Kohl Alexandre Djandji Stephanie Morgan Susan Whittier Mahesh Mansukhani Eldad Hod Mary D’Alton Yousin Suh Zev Williams |
| author_sort |
Shan Wei |
| title |
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| title_short |
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| title_full |
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| title_fullStr |
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| title_full_unstemmed |
Direct diagnostic testing of SARS-CoV-2 without the need for prior RNA extraction |
| title_sort |
direct diagnostic testing of sars-cov-2 without the need for prior rna extraction |
| publisher |
Nature Portfolio |
| publishDate |
2021 |
| url |
https://doaj.org/article/6d182b455806472a8759a067b2e4669f |
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