RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)

BACKGROUND: Current diagnosis of SARS-CoV-2 infection relies on RNA purification prior to amplification. Typical extraction methods limit the processing speed and turnaround time for SARS-CoV-2 diagnostic testing. METHODS: Here, we applied reverse transcription loop-mediated isothermal amplification...

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Autores principales: Lai, Meng Yee, Suppiah, Jeyanthi, Thayan, Ravindran, Ismail, Ilyiana, Mustapa, Nur Izati, Soh, Tuan Suhaila Tuan, Hassan, Afifah Haji, Peariasamy, Kalaiarasu M., Lee, Yee Leng, Lau, Yee Ling
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Short Report
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8723997/
https://www.ncbi.nlm.nih.gov/pubmed/34980275
http://dx.doi.org/10.1186/s41182-021-00396-y
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author Lai, Meng Yee
Suppiah, Jeyanthi
Thayan, Ravindran
Ismail, Ilyiana
Mustapa, Nur Izati
Soh, Tuan Suhaila Tuan
Hassan, Afifah Haji
Peariasamy, Kalaiarasu M.
Lee, Yee Leng
Lau, Yee Ling
author_facet Lai, Meng Yee
Suppiah, Jeyanthi
Thayan, Ravindran
Ismail, Ilyiana
Mustapa, Nur Izati
Soh, Tuan Suhaila Tuan
Hassan, Afifah Haji
Peariasamy, Kalaiarasu M.
Lee, Yee Leng
Lau, Yee Ling
author_sort Lai, Meng Yee
collection PubMed
description BACKGROUND: Current diagnosis of SARS-CoV-2 infection relies on RNA purification prior to amplification. Typical extraction methods limit the processing speed and turnaround time for SARS-CoV-2 diagnostic testing. METHODS: Here, we applied reverse transcription loop-mediated isothermal amplification directly onto human clinical swabs samples to amplify the RNA from SARS-CoV-2 swab samples after processing with chelating resin. RESULTS: By testing our method on 64 samples, we managed to develop an RT-LAMP assay with 95.9% sensitivity (95% CI 86 to 99.5%) and 100% specificity (95% CI 78.2–100%). CONCLUSION: The entire process including sample processing can be completed in approximately 50 min. This method has promising potential to be applied as a fast, simple and inexpensive diagnostic tool for the detection of SARS-CoV-2. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s41182-021-00396-y.
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spelling pubmed-87239972022-01-04 RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP) Lai, Meng Yee Suppiah, Jeyanthi Thayan, Ravindran Ismail, Ilyiana Mustapa, Nur Izati Soh, Tuan Suhaila Tuan Hassan, Afifah Haji Peariasamy, Kalaiarasu M. Lee, Yee Leng Lau, Yee Ling Trop Med Health Short Report BACKGROUND: Current diagnosis of SARS-CoV-2 infection relies on RNA purification prior to amplification. Typical extraction methods limit the processing speed and turnaround time for SARS-CoV-2 diagnostic testing. METHODS: Here, we applied reverse transcription loop-mediated isothermal amplification directly onto human clinical swabs samples to amplify the RNA from SARS-CoV-2 swab samples after processing with chelating resin. RESULTS: By testing our method on 64 samples, we managed to develop an RT-LAMP assay with 95.9% sensitivity (95% CI 86 to 99.5%) and 100% specificity (95% CI 78.2–100%). CONCLUSION: The entire process including sample processing can be completed in approximately 50 min. This method has promising potential to be applied as a fast, simple and inexpensive diagnostic tool for the detection of SARS-CoV-2. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s41182-021-00396-y. BioMed Central 2022-01-04 /pmc/articles/PMC8723997/ /pubmed/34980275 http://dx.doi.org/10.1186/s41182-021-00396-y Text en © The Author(s) 2022, corrected publication 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Short Report
Lai, Meng Yee
Suppiah, Jeyanthi
Thayan, Ravindran
Ismail, Ilyiana
Mustapa, Nur Izati
Soh, Tuan Suhaila Tuan
Hassan, Afifah Haji
Peariasamy, Kalaiarasu M.
Lee, Yee Leng
Lau, Yee Ling
RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_full RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_fullStr RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_full_unstemmed RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_short RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
title_sort rna purification-free detection of sars-cov-2 using reverse transcription loop-mediated isothermal amplification (rt-lamp)
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8723997/
https://www.ncbi.nlm.nih.gov/pubmed/34980275
http://dx.doi.org/10.1186/s41182-021-00396-y
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