Evaluation of two lyophilized molecular assays to rapidly detect foot-and-mouth disease virus directly from clinical samples in field settings

Howson, E.L.A. et al. (2017) Evaluation of two lyophilized molecular assays to rapidly detect foot-and-mouth disease virus directly from clinical samples in field settings. Transboundary and Emerging Diseases, 64(3), pp. 861-871. (doi:10.1111/tbed.12451) (PMID:26617330) (PMCID:PMC5434942)

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Abstract

Accurate, timely diagnosis is essential for the control, monitoring and eradication of foot‐and‐mouth disease (FMD). Clinical samples from suspect cases are normally tested at reference laboratories. However, transport of samples to these centralized facilities can be a lengthy process that can impose delays on critical decision making. These concerns have motivated work to evaluate simple‐to‐use technologies, including molecular‐based diagnostic platforms, that can be deployed closer to suspect cases of FMD. In this context, FMD virus (FMDV)‐specific reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) and real‐time RT‐PCR (rRT‐PCR) assays, compatible with simple sample preparation methods and in situ visualization, have been developed which share equivalent analytical sensitivity with laboratory‐based rRT‐PCR. However, the lack of robust ‘ready‐to‐use kits’ that utilize stabilized reagents limits the deployment of these tests into field settings. To address this gap, this study describes the performance of lyophilized rRT‐PCR and RT‐LAMP assays to detect FMDV. Both of these assays are compatible with the use of fluorescence to monitor amplification in real‐time, and for the RT‐LAMP assays end point detection could also be achieved using molecular lateral flow devices. Lyophilization of reagents did not adversely affect the performance of the assays. Importantly, when these assays were deployed into challenging laboratory and field settings within East Africa they proved to be reliable in their ability to detect FMDV in a range of clinical samples from acutely infected as well as convalescent cattle. These data support the use of highly sensitive molecular assays into field settings for simple and rapid detection of FMDV.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Lembo, Dr Tiziana and Cleaveland, Professor Sarah and Howson, Emma and Armson, Bryony
Authors: Howson, E.L.A., Armson, B., Madi, M., Kasanga, C.J., Kandusi, S., Sallu, R., Chepkwony, E., Siddle, A., Martin, P., Wood, J., Mioulet, V., King, D.P., Lembo, T., Cleaveland, S., and Fowler, V.L.
College/School:College of Medical Veterinary and Life Sciences > Institute of Biodiversity Animal Health and Comparative Medicine
Journal Name:Transboundary and Emerging Diseases
Publisher:Wiley
ISSN:1865-1674
ISSN (Online):1865-1682
Published Online:30 November 2015
Copyright Holders:Copyright © 2015 The Authors
First Published:First published in Transboundary and Emerging Diseases 64(3): 861-871
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
518811Towards the strategic control of foot-and-mouth disease in Africa: new techniques for a neglected problemSarah CleavelandBiotechnology and Biological Sciences Research Council (BBSRC)BB/H009302/1RI BIODIVERSITY ANIMAL HEALTH & COMPMED