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A flexible format LAMP assay for rapid detection of Ebola virus

BACKGROUND: The unprecedented 2013/16 outbreak of Zaire ebolavirus (Ebola virus) in West Africa has highighted the need for rapid, high-throughput and POC diagnostic assays to enable timely detection and appropriate triaging of Ebola Virus Disease (EVD) patients. Ebola virus is highly infectious and...

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Autores principales: Bonney, Laura C., Watson, Robert J., Slack, Gillian S., Bosworth, Andrew, Wand, Nadina I. Vasileva, Hewson, Roger
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7423149/
https://www.ncbi.nlm.nih.gov/pubmed/32735587
http://dx.doi.org/10.1371/journal.pntd.0008496
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author Bonney, Laura C.
Watson, Robert J.
Slack, Gillian S.
Bosworth, Andrew
Wand, Nadina I. Vasileva
Hewson, Roger
author_facet Bonney, Laura C.
Watson, Robert J.
Slack, Gillian S.
Bosworth, Andrew
Wand, Nadina I. Vasileva
Hewson, Roger
author_sort Bonney, Laura C.
collection PubMed
description BACKGROUND: The unprecedented 2013/16 outbreak of Zaire ebolavirus (Ebola virus) in West Africa has highighted the need for rapid, high-throughput and POC diagnostic assays to enable timely detection and appropriate triaging of Ebola Virus Disease (EVD) patients. Ebola virus is highly infectious and prompt diagnosis and triage is crucial in preventing further spread within community and healthcare settings. Moreover, due to the ecology of Ebola virus it is important that newly developed diagnostic assays are suitable for use in both the healthcare environment and low resource rural locations. METHODOLOGY/PRINCIPLE FINDINGS: A LAMP assay was successfully developed with three detection formats; a real-time intercalating dye-based assay, a real-time probe-based assay to enable multiplexing and an end-point colourimetric assay to simplify interpretation for the field. All assay formats were sensitive and specific, detecting a range of Ebola virus strains isolated in 1976–2014; with Probit analysis predicting limits of detection of 243, 290 and 75 copies/reaction respectively and no cross-detection of related strains or other viral haemorrhagic fevers (VHF’s). The assays are rapid, (as fast as 5–7.25 mins for real-time formats) and robust, detecting Ebola virus RNA in presence of minimally diluted bodily fluids. Moreover, when tested on patient samples from the 2013/16 outbreak, there were no false positives and 93–96% of all new case positives were detected, with only a failure to detect very low copy number samples. CONCLUSION/SIGNIFICANCE: These are a set of robust and adaptable diagnostic solutions, which are fast, easy-to-perform-and-interpret and are suitable for use on a range of platforms including portable low-power devices. They can be readily transferred to field-laboratory settings, with no specific equipment needs and are therefore ideally placed for use in locations with limited resources.
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spelling pubmed-74231492020-08-20 A flexible format LAMP assay for rapid detection of Ebola virus Bonney, Laura C. Watson, Robert J. Slack, Gillian S. Bosworth, Andrew Wand, Nadina I. Vasileva Hewson, Roger PLoS Negl Trop Dis Research Article BACKGROUND: The unprecedented 2013/16 outbreak of Zaire ebolavirus (Ebola virus) in West Africa has highighted the need for rapid, high-throughput and POC diagnostic assays to enable timely detection and appropriate triaging of Ebola Virus Disease (EVD) patients. Ebola virus is highly infectious and prompt diagnosis and triage is crucial in preventing further spread within community and healthcare settings. Moreover, due to the ecology of Ebola virus it is important that newly developed diagnostic assays are suitable for use in both the healthcare environment and low resource rural locations. METHODOLOGY/PRINCIPLE FINDINGS: A LAMP assay was successfully developed with three detection formats; a real-time intercalating dye-based assay, a real-time probe-based assay to enable multiplexing and an end-point colourimetric assay to simplify interpretation for the field. All assay formats were sensitive and specific, detecting a range of Ebola virus strains isolated in 1976–2014; with Probit analysis predicting limits of detection of 243, 290 and 75 copies/reaction respectively and no cross-detection of related strains or other viral haemorrhagic fevers (VHF’s). The assays are rapid, (as fast as 5–7.25 mins for real-time formats) and robust, detecting Ebola virus RNA in presence of minimally diluted bodily fluids. Moreover, when tested on patient samples from the 2013/16 outbreak, there were no false positives and 93–96% of all new case positives were detected, with only a failure to detect very low copy number samples. CONCLUSION/SIGNIFICANCE: These are a set of robust and adaptable diagnostic solutions, which are fast, easy-to-perform-and-interpret and are suitable for use on a range of platforms including portable low-power devices. They can be readily transferred to field-laboratory settings, with no specific equipment needs and are therefore ideally placed for use in locations with limited resources. Public Library of Science 2020-07-31 /pmc/articles/PMC7423149/ /pubmed/32735587 http://dx.doi.org/10.1371/journal.pntd.0008496 Text en © 2020 Bonney et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Bonney, Laura C.
Watson, Robert J.
Slack, Gillian S.
Bosworth, Andrew
Wand, Nadina I. Vasileva
Hewson, Roger
A flexible format LAMP assay for rapid detection of Ebola virus
title A flexible format LAMP assay for rapid detection of Ebola virus
title_full A flexible format LAMP assay for rapid detection of Ebola virus
title_fullStr A flexible format LAMP assay for rapid detection of Ebola virus
title_full_unstemmed A flexible format LAMP assay for rapid detection of Ebola virus
title_short A flexible format LAMP assay for rapid detection of Ebola virus
title_sort flexible format lamp assay for rapid detection of ebola virus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7423149/
https://www.ncbi.nlm.nih.gov/pubmed/32735587
http://dx.doi.org/10.1371/journal.pntd.0008496
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