title: Recovery of viral RNA and infectious foot-and-mouth disease virus from positive lateral-flow devices.
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.
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Veronica L
authors.initials: VL
authors.lastname: Fowler
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Bartlomiej M
authors.initials: BM
authors.lastname: Bankowski
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Bryony
authors.initials: B
authors.lastname: Armson
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom; Institute of Biodiversity, Animal Health and Comparative Medicine, College of Medical, Veterinary and Life Science, University of Glasgow, Glasgow, United Kingdom.
authors.forename: Antonello
authors.initials: A
authors.lastname: Di Nardo
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Begoña
authors.initials: B
authors.lastname: Valdazo-Gonzalez
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom; Animal Health and Veterinary Laboratories Agency (AHVLA), New Haw, Surrey, United Kingdom.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Reid
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom; Veterinary Medicines Directorate, New Haw, Surrey, United Kingdom.
authors.forename: Paul V
authors.initials: PV
authors.lastname: Barnett
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Jemma
authors.initials: J
authors.lastname: Wadsworth
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Nigel P
authors.initials: NP
authors.lastname: Ferris
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Valérie
authors.initials: V
authors.lastname: Mioulet
authors.affiliation: Vesicular Disease Reference Laboratory, The Pirbright Institute, Pirbright, Surrey, United Kingdom.
authors.forename: Donald P
authors.initials: DP
authors.lastname: King
mesh_terms: D000818:Animals; D002460:Cell Line; D005536:Foot-and-Mouth Disease / Q000473:pathology / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics* / Q000302:isolation & purification; D016679:Genome, Viral; D006168:Guinea Pigs; D006801:Humans; D012367:RNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D012703:Serotyping; D013048:Specimen Handling / Q000295:instrumentation*; D013696:Temperature
chemical_list: D012367:RNA, Viral