Detection of capripoxvirus DNA using a field-ready nucleic acid extraction and real-time PCR platform

Armson, B. et al. (2015) Detection of capripoxvirus DNA using a field-ready nucleic acid extraction and real-time PCR platform. Transboundary and Emerging Diseases, 64(3), pp. 994-997. (doi:10.1111/tbed.12447) (PMID:26608662)

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Abstract

Capripoxviruses, comprising sheep pox virus, goat pox virus and lumpy skin disease virus cause serious diseases of domesticated ruminants, notifiable to The World Organization for Animal Health. This report describes the evaluation of a mobile diagnostic system (Enigma Field Laboratory) that performs automated sequential steps for nucleic acid extraction and real-time PCR to detect capripoxvirus DNA within laboratory and endemic field settings. To prepare stable reagents that could be deployed into field settings, lyophilized reagents were used that employed an established diagnostic PCR assay. These stabilized reagents demonstrated an analytical sensitivity that was equivalent, or greater than the established laboratory-based PCR test which utilizes wet reagents, and the limit of detection for the complete assay pipeline was approximately one log10 more sensitive than the laboratory-based PCR assay. Concordant results were generated when the mobile PCR system was compared to the laboratory-based PCR using samples collected from Africa, Asia and Europe (n = 10) and experimental studies (n = 9) representing clinical cases of sheep pox, goat pox and lumpy skin disease. Furthermore, this mobile assay reported positive results in situ using specimens that were collected from a dairy cow in Morogoro, Tanzania, which was exhibiting clinical signs of lumpy skin disease. These data support the use of mobile PCR systems for the rapid and sensitive detection of capripoxvirus DNA in endemic field settings.

Item Type:Articles
Additional Information:The authors acknowledge the support provided from the European Union FP7-KBBE-2011-5 under grant agreement no. 289364 (RAPIDIA-Field), The Wellcome Trust via funding for the Southern African Centre for Disease Surveillance (SACIDS) grant WT087546MA and the Department of Environment, Food and Rural Affairs for supporting the work of the Non-Vesicular Reference Laboratory at The Pirbright Institute. Additional financial support for field work in Tanzania was provided via a one year research grant from EuFMD-FAR funded by the European Union.
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:UNSPECIFIED
Authors: Armson, B., Fowler, V. L., Tuppurainen, E. S. M., Howson, E.L.A., Madi, M., Sallu, R., Kasanga, C. J., Pearson, C., Wood, J., Martin, P., Mioulet, V., and King, D. P.
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:25 November 2015
Copyright Holders:Copyright © 2017 The Authors
First Published:First published in Transboundary and Emerging Diseases 64(3):994-997
Publisher Policy:Reproduced under a Creative Commons License

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