The development of two field‐ready reverse transcription loop‐mediated isothermal amplification assays for the rapid detection of Seneca Valley virus 1

Armson, B., Walsh, C., Morant, N., Fowler, V. L., Knowles, N. J. and Clark, D. (2018) The development of two field‐ready reverse transcription loop‐mediated isothermal amplification assays for the rapid detection of Seneca Valley virus 1. Transboundary and Emerging Diseases, 66(1), pp. 497-504. (doi: 10.1111/tbed.13051) (PMID:30372584) (PMCID:PMC6434928)

Abstract

Seneca Valley virus 1 (SVV‐1) has been associated with vesicular disease in swine, with clinical signs indistinguishable from those of other notifiable vesicular diseases such as foot‐and‐mouth disease. Rapid and accurate detection of SVV‐1 is central to confirm the disease causing agent, and to initiate the implementation of control processes. The development of rapid, cost‐effective diagnostic assays that can be used at the point of sample collection has been identified as a gap in preparedness for the control of SVV‐1. This study describes the development and bench validation of two reverse transcription loop‐mediated amplification (RT‐LAMP) assays targeting the 5′‐untranslated region (5′‐UTR) and the VP3‐1 region for the detection of SVV‐1 that may be performed at the point of sample collection. Both assays were able to demonstrate amplification of all neat samples diluted 1/100 in negative pig epithelium tissue suspension within 8 min, when RNA was extracted prior to the RT‐LAMP assay, and no amplification was observed for the other viruses tested. Simple sample preparation methods using lyophilized reagents were investigated, to negate the requirement for RNA extraction. Only a small delay in the time to amplification was observed for these lyophilized reagents, with a time from sample receipt to amplification achieved within 12 min. Although diagnostic validation is recommended, these RT‐LAMP assays are highly sensitive and specific, with the potential to be a useful tool in the rapid diagnosis of SVV‐1 in the field.

• Item Type: Articles
• Additional Information: This study was carried out as part of an industrial placement on a BBSRC CASE PhD studentship 1646343, related to BBSRC grant number BB/M017273/1, for which the authors acknowledge.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Fowler, Dr Veronica and Armson, Bryony
• Authors: Armson, B., Walsh, C., Morant, N., Fowler, V. L., Knowles, N. J., and Clark, D.
• College/School: College of Medical Veterinary and Life Sciences > School of Biodiversity, One Health & Veterinary Medicine
• Journal Name: Transboundary and Emerging Diseases
• Publisher: Wiley
• ISSN: 1865-1674
• ISSN (Online): 1865-1682
• Published Online: 29 October 2018
• Copyright Holders: Copyright © 2018 GeneSys Biotech Limited/OptiGene Limited
• First Published: First published in Transboundary and Emerging Diseases 66(1):497-504
• Publisher Policy: Reproduced under a Creative Commons License