Recovery of viral RNA and infectious foot-and-mouth disease virus from positive lateral-flow devices

PLoS One. 2014 Oct 14;9(10):e109322. doi: 10.1371/journal.pone.0109322. eCollection 2014.

Abstract

Foot-and-mouth disease Virus (FMDV) is an economically important, highly contagious picornavirus that affects both wild and domesticated cloven hooved animals. In developing countries, the effective laboratory diagnosis of foot-and-mouth disease (FMD) is often hindered by inadequate sample preservation due to difficulties in the transportation and storage of clinical material. These factors can compromise the ability to detect and characterise FMD virus in countries where the disease is endemic. Furthermore, the high cost of sending infectious virus material and the biosecurity risk it presents emphasises the need for a thermo-stable, non-infectious mode of transporting diagnostic samples. This paper investigates the potential of using FMDV lateral-flow devices (LFDs) for dry transportation of clinical samples for subsequent nucleic acid amplification, sequencing and recovery of infectious virus by electroporation. FMDV positive samples (epithelial suspensions and cell culture isolates) representing four FMDV serotypes were applied to antigen LFDs: after which it was possible to recover viral RNA that could be detected using real-time RT-PCR. Using this nucleic acid, it was also possible to recover VP1 sequences and also successfully utilise protocols for amplification of complete FMD virus genomes. It was not possible to recover infectious FMDV directly from the LFDs, however following electroporation into BHK-21 cells and subsequent cell passage, infectious virus could be recovered. Therefore, these results support the use of the antigen LFD for the dry, non-hazardous transportation of samples from FMD endemic countries to international reference laboratories.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Foot-and-Mouth Disease / pathology
  • Foot-and-Mouth Disease / virology*
  • Foot-and-Mouth Disease Virus / genetics*
  • Foot-and-Mouth Disease Virus / isolation & purification
  • Genome, Viral
  • Guinea Pigs
  • Humans
  • RNA, Viral / analysis*
  • RNA, Viral / isolation & purification
  • Real-Time Polymerase Chain Reaction
  • Sequence Analysis, RNA
  • Serotyping
  • Specimen Handling / instrumentation*
  • Temperature

Substances

  • RNA, Viral

Grants and funding

The work was supported financially by Department for the Environment, Food and Rural Affairs (DEFRA SE1127) and funding from the European Union FP7-KBBE-2011-5 under grant agreement no. 289364 (RAPIDIA-Field). In addition this material is based upon work supported by the U.S. Department of Homeland Security under Award Number HSHQDC-12-J-00415 through Texas A&M AgriLife Research and the Institute for Infectious Diseases (IIAD). The views and conclusions contained in this document are those of the authors and should not be interpreted as necessarily representing the official policies, either expressed or implied, of the U.S. Department of Homeland Security. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.