A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens
Abstract Access to fast and reliable nucleic acid testing continues to play a key role in controlling the COVID-19 pandemic, especially in the context of increased vaccine break-through risks due to new variants. We report a rapid, low-cost (~ 2 USD), simple-to-use nucleic acid test kit for self-adm...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2022-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-022-11144-5 |
| _version_ | 1811289408225148928 |
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| author | Xin Song Felicity J. Coulter Ming Yang Jessica L. Smith Fikadu G. Tafesse William B. Messer John H. Reif |
| author_facet | Xin Song Felicity J. Coulter Ming Yang Jessica L. Smith Fikadu G. Tafesse William B. Messer John H. Reif |
| author_sort | Xin Song |
| collection | DOAJ |
| description | Abstract Access to fast and reliable nucleic acid testing continues to play a key role in controlling the COVID-19 pandemic, especially in the context of increased vaccine break-through risks due to new variants. We report a rapid, low-cost (~ 2 USD), simple-to-use nucleic acid test kit for self-administered at-home testing without lab instrumentation. The entire sample-to-answer workflow takes < 60 min, including noninvasive sample collection, one-step RNA preparation, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) in a thermos, and direct visual inspection of a colorimetric test result. To facilitate long-term storage without cold-chain, a fast one-pot lyophilization protocol was developed to preserve all required biochemical reagents of the colorimetric RT-LAMP test in a single microtube. Notably, the lyophilized RT-LAMP assay demonstrated reduced false positives as well as enhanced tolerance to a wider range of incubation temperatures compared to solution-based RT-LAMP reactions. We validated our RT-LAMP assay using simulated infected samples, and detected a panel of SARS-CoV-2 variants with successful detection of all variants that were available to us at the time. With a simple change of the primer set, our lyophilized RT-LAMP home test can be easily adapted as a low-cost surveillance platform for other pathogens and infectious diseases of global public health importance. |
| first_indexed | 2024-04-13T03:54:31Z |
| format | Article |
| id | doaj.art-404aec15e5864b4da7c4cf9f4bbc274e |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-13T03:54:31Z |
| publishDate | 2022-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-404aec15e5864b4da7c4cf9f4bbc274e2022-12-22T03:03:41ZengNature PortfolioScientific Reports2045-23222022-04-0112111110.1038/s41598-022-11144-5A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogensXin Song0Felicity J. Coulter1Ming Yang2Jessica L. Smith3Fikadu G. Tafesse4William B. Messer5John H. Reif6Department of Electrical and Computer Engineering, Duke UniversityDepartment of Molecular Microbiology and Immunology, Oregon Health and Science UniversityDepartment of Computer Science, Duke UniversityVaccine and Gene Therapy Institute, Oregon Health and Science UniversityDepartment of Molecular Microbiology and Immunology, Oregon Health and Science UniversityDepartment of Molecular Microbiology and Immunology, Oregon Health and Science UniversityDepartment of Electrical and Computer Engineering, Duke UniversityAbstract Access to fast and reliable nucleic acid testing continues to play a key role in controlling the COVID-19 pandemic, especially in the context of increased vaccine break-through risks due to new variants. We report a rapid, low-cost (~ 2 USD), simple-to-use nucleic acid test kit for self-administered at-home testing without lab instrumentation. The entire sample-to-answer workflow takes < 60 min, including noninvasive sample collection, one-step RNA preparation, reverse-transcription loop-mediated isothermal amplification (RT-LAMP) in a thermos, and direct visual inspection of a colorimetric test result. To facilitate long-term storage without cold-chain, a fast one-pot lyophilization protocol was developed to preserve all required biochemical reagents of the colorimetric RT-LAMP test in a single microtube. Notably, the lyophilized RT-LAMP assay demonstrated reduced false positives as well as enhanced tolerance to a wider range of incubation temperatures compared to solution-based RT-LAMP reactions. We validated our RT-LAMP assay using simulated infected samples, and detected a panel of SARS-CoV-2 variants with successful detection of all variants that were available to us at the time. With a simple change of the primer set, our lyophilized RT-LAMP home test can be easily adapted as a low-cost surveillance platform for other pathogens and infectious diseases of global public health importance.https://doi.org/10.1038/s41598-022-11144-5 |
| spellingShingle | Xin Song Felicity J. Coulter Ming Yang Jessica L. Smith Fikadu G. Tafesse William B. Messer John H. Reif A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens Scientific Reports |
| title | A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens |
| title_full | A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens |
| title_fullStr | A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens |
| title_full_unstemmed | A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens |
| title_short | A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens |
| title_sort | lyophilized colorimetric rt lamp test kit for rapid low cost at home molecular testing of sars cov 2 and other pathogens |
| url | https://doi.org/10.1038/s41598-022-11144-5 |
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