A lyophilized colorimetric RT-LAMP test kit for rapid, low-cost, at-home molecular testing of SARS-CoV-2 and other pathogens.

Autor: Song X; Department of Electrical and Computer Engineering, Duke University, Durham, NC, 27708, USA. xin.song@duke.edu.; Department of Biomedical Engineering, Duke University, Durham, NC, 27708, USA. xin.song@duke.edu.; Department of Computer Science, Duke University, Durham, NC, 27708, USA. xin.song@duke.edu., Coulter FJ; Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR, 97239, USA., Yang M; Department of Computer Science, Duke University, Durham, NC, 27708, USA., Smith JL; Vaccine and Gene Therapy Institute, Oregon Health and Science University, Beaverton, OR, 97006, USA., Tafesse FG; Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR, 97239, USA., Messer WB; Department of Molecular Microbiology and Immunology, Oregon Health and Science University, Portland, OR, 97239, USA. messer@ohsu.edu.; Department of Medicine, Division of Infectious Diseases, Oregon Health and Science University, Portland, OR, 97239, USA. messer@ohsu.edu.; Program in Epidemiology, OHSU-PSU School of Public Health, Oregon Health and Science University, Portland, OR, 97239, USA. messer@ohsu.edu., Reif JH; Department of Electrical and Computer Engineering, Duke University, Durham, NC, 27708, USA. reif@cs.duke.edu.; Department of Computer Science, Duke University, Durham, NC, 27708, USA. reif@cs.duke.edu.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2022 Apr 29; Vol. 12 (1), pp. 7043. Date of Electronic Publication: 2022 Apr 29.
DOI: 10.1038/s41598-022-11144-5
Abstrakt: 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.
(© 2022. The Author(s).)
Databáze: MEDLINE
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