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...

Descripción completa

Detalles Bibliográficos
Autores principales: 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
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