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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Virology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1429288

Detection of foot-and-mouth disease virus RNA using a closed loopmediated isothermal amplification system

Provisionally accepted
Natasha Edwards Natasha Edwards 1Julien Reboud Julien Reboud 2Xiaoxiang Yan Xiaoxiang Yan 2Xin Guo Xin Guo 2Jonathan M. Cooper Jonathan M. Cooper 2Jemma Wadsworth Jemma Wadsworth 1Ryan Waters Ryan Waters 1Valerie Mioulet Valerie Mioulet 1Donald King Donald King 1Andrew E. Shaw Andrew E. Shaw 1*
  • 1 The Pirbright Institute, Woking, England, United Kingdom
  • 2 University of Glasgow, Glasgow, Scotland, United Kingdom

The final, formatted version of the article will be published soon.

    Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals responsible for economic losses that amount to >$20 billion annually. Rapid recognition of FMD cases provides vital information to guide control programs. A range of point-of-need (PON) amplification technologies have been developed which allow sensitive detection of the causative virus (FMDV) in the field at locations remote from laboratories. Here we describe a novel system to detect FMDV RNA using loop mediated isothermal amplification (LAMP). This test was evaluated using a panel of FMDV isolates (n=79) and RNA standards demonstrating capability to amplify viral genome directly from clinical material in the absence of nucleic acid extraction. This extraction-free RT-LAMP assay was transferred to a bespoke closed-system lateral flow test (LFT) that was used in combination with a lowcost hand-held heater. Our results show that the RT-LAMP-LFT assay retains a high level of diagnostic and analytical sensitivity when using direct clinical material, with a limit of detection of approximately 80 copies per reaction. Together, our data support the potential for the use of this assay at the PON to facilitate rapid feedback on the status of suspect cases.

    Keywords: Foot-and-Mouth Disease, virus, Loop-mediated amplification, LAMP, point-of-care, diagnosis, bioengineering Clinical samples were collected using

    Received: 07 May 2024; Accepted: 18 Jul 2024.

    Copyright: © 2024 Edwards, Reboud, Yan, Guo, Cooper, Wadsworth, Waters, Mioulet, King and Shaw. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Andrew E. Shaw, The Pirbright Institute, Woking, GU24 0NF, England, United Kingdom

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.