A flexible format lamp assay for rapid detection of ebola virus

Laura C. Bonney*, Robert J. Watson, Gillian S. Slack, Andrew Bosworth, Nadina I. Vasileva Wand, Roger Hewson

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

5 Citations (Scopus)
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Abstract

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 interca-lating 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-per-form-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.

Original languageEnglish
Article numbere0008496
Pages (from-to)1-22
Number of pages22
JournalPLoS Neglected Tropical Diseases
Volume14
Issue number7
DOIs
Publication statusPublished - 31 Jul 2020

Bibliographical note

Funding Information: This project has received funding from the Innovative Medicines Initiative 2 Joint Undertaking under grant agreement No 115843. This Joint Undertaking receives support from the European Union's Horizon 2020 research and innovation programme and EFPIA. (www.imi. europa.eu) Funding awarded to Professor Roger Hewson as part of the EbolaMoDRAD programme. To the best of my knowledge the funders did not play any role in the study design, data collection and analysis, decision to publish or preparation of the manuscript.

Open Access: This is an open access article distributed under the terms of the
Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publisher Copyright:© 2020 Bonney et al.

Citation: Bonney LC, Watson RJ, Slack GS, Bosworth A, Wand NIV, Hewson R (2020) A flexible format LAMP assay for rapid detection of Ebola virus. PLoS Negl Trop Dis 14(7): e0008496.

DOI: https://doi.org/10.1371/journal.pntd.0008496

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