Cargando…
Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2
BACKGROUND: Current assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595270/ https://www.ncbi.nlm.nih.gov/pubmed/34789179 http://dx.doi.org/10.1186/s12879-021-06876-0 |
_version_ | 1784600163688382464 |
---|---|
author | Lai, Meng Yee Bukhari, Fatma Diyana Mohd Zulkefli, Nur Zulaikha Ismail, Ilyiana Mustapa, Nur Izati Soh, Tuan Suhaila Tuan Hassan, Afifah Haji Peariasamy, Kalaiarasu M. Lee, Yee Leng Suppiah, Jeyanthi Thayan, Ravindran Lau, Yee Ling |
author_facet | Lai, Meng Yee Bukhari, Fatma Diyana Mohd Zulkefli, Nur Zulaikha Ismail, Ilyiana Mustapa, Nur Izati Soh, Tuan Suhaila Tuan Hassan, Afifah Haji Peariasamy, Kalaiarasu M. Lee, Yee Leng Suppiah, Jeyanthi Thayan, Ravindran Lau, Yee Ling |
author_sort | Lai, Meng Yee |
collection | PubMed |
description | BACKGROUND: Current assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from nasopharyngeal swab samples that bypasses the RNA purification step. METHODS: In the current project, we have described two extraction-free reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of SARS-CoV-2 by using E gene and RdRp gene as the targets. RESULTS: Here, results showed that reverse transcription loop-mediated isothermal amplification assays with 88.4% sensitive (95% CI: 74.9–96.1%) and 67.4% sensitive (95% CI: 51.5–80.9%) for E gene and RdRp gene, respectively. CONCLUSION: Without the need of RNA purification, our developed RT-LAMP assays for direct detection of SARS-CoV-2 from nasopharyngeal swab samples could be turned into alternatives to qRT-PCR for rapid screening. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-021-06876-0. |
format | Online Article Text |
id | pubmed-8595270 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-85952702021-11-17 Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 Lai, Meng Yee Bukhari, Fatma Diyana Mohd Zulkefli, Nur Zulaikha Ismail, Ilyiana Mustapa, Nur Izati Soh, Tuan Suhaila Tuan Hassan, Afifah Haji Peariasamy, Kalaiarasu M. Lee, Yee Leng Suppiah, Jeyanthi Thayan, Ravindran Lau, Yee Ling BMC Infect Dis Research BACKGROUND: Current assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from nasopharyngeal swab samples that bypasses the RNA purification step. METHODS: In the current project, we have described two extraction-free reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of SARS-CoV-2 by using E gene and RdRp gene as the targets. RESULTS: Here, results showed that reverse transcription loop-mediated isothermal amplification assays with 88.4% sensitive (95% CI: 74.9–96.1%) and 67.4% sensitive (95% CI: 51.5–80.9%) for E gene and RdRp gene, respectively. CONCLUSION: Without the need of RNA purification, our developed RT-LAMP assays for direct detection of SARS-CoV-2 from nasopharyngeal swab samples could be turned into alternatives to qRT-PCR for rapid screening. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12879-021-06876-0. BioMed Central 2021-11-17 /pmc/articles/PMC8595270/ /pubmed/34789179 http://dx.doi.org/10.1186/s12879-021-06876-0 Text en © The Author(s) 2021, 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/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
spellingShingle | Research Lai, Meng Yee Bukhari, Fatma Diyana Mohd Zulkefli, Nur Zulaikha Ismail, Ilyiana Mustapa, Nur Izati Soh, Tuan Suhaila Tuan Hassan, Afifah Haji Peariasamy, Kalaiarasu M. Lee, Yee Leng Suppiah, Jeyanthi Thayan, Ravindran Lau, Yee Ling Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title | Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title_full | Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title_fullStr | Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title_full_unstemmed | Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title_short | Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2 |
title_sort | two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of sars-cov-2 |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8595270/ https://www.ncbi.nlm.nih.gov/pubmed/34789179 http://dx.doi.org/10.1186/s12879-021-06876-0 |
work_keys_str_mv | AT laimengyee twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT bukharifatmadiyanamohd twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT zulkeflinurzulaikha twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT ismaililyiana twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT mustapanurizati twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT sohtuansuhailatuan twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT hassanafifahhaji twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT peariasamykalaiarasum twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT leeyeeleng twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT suppiahjeyanthi twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT thayanravindran twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 AT lauyeeling twoextractionfreereversetranscriptionloopmediatedisothermalamplificationassaysfordetectionofsarscov2 |