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  • Author: Bachanek-Bankowska, Katarzyna

    The Pirbright Institute, United Kingdom

  • Author: Mero, Herieth R.

    Sokoine University of Agriculture, Tanzania, United Republic of

  • Author: Wadsworth, Jemma

    The Pirbright Institute, United Kingdom

  • Author: Mioulet, Valerie

    The Pirbright Institute, United Kingdom

  • Author: Sallu, Raphael

    Tanzania Veterinary Laboratory Agency, Tanzania, United Republic of

  • Author: Belsham, Graham

    Section for Virology, National Veterinary Institute, Technical University of Denmark, Lindholm, 4771, Kalvehave, Denmark

  • Author: Kasanga, Christopher J.

    Sokoine University of Agriculture, Tanzania, United Republic of

  • Author: Knowles, Nick J.

    The Pirbright Institute, United Kingdom

  • Author: King, Donald P.

    The Pirbright Institute, United Kingdom

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Rapid, reliable and accurate diagnostic methods provide essential support to programmes that monitor and control foot-and-mouth disease (FMD). While pan-specific molecular tests for FMD virus (FMDV) detection are well established and widely used in endemic and FMD-free countries, current serotyping methods mainly rely either on antigen detection ELISAs or nucleotide sequencing approaches. This report describes the development of a panel of serotype-specific real-time RT-PCR assays (rRT-PCR) tailored to detect FMDV lineages currently circulating in East Africa. These assays target sequences within the VP1-coding region that share high intra-lineage identity, but do not cross-react with FMD viruses from other serotypes that circulate in the region. These serotype-specific assays operate with the same thermal profile as the pan-diagnostic tests making it possible to run them in parallel to produce CT values comparable to the pan-diagnostic test detecting the 3D-coding region. These assays were evaluated alongside the established pan-specific molecular test using field samples and virus isolates collected from Tanzania, Kenya and Ethiopia that had been previously characterised by nucleotide sequencing. Samples (n = 71) representing serotype A (topotype AFRICA, lineage G-I), serotype O (topotypes EA-2 and EA-4), serotype SAT 1 (topotype I (NWZ)) and serotype SAT2 (topotype IV) were correctly identified with these rRT-PCR assays. Furthermore, FMDV RNA from samples that did not contain infectious virus could still be serotyped using these assays. These serotype-specific real-time RT-PCR assays can detect and characterise FMDVs currently circulating in East Africa and hence improve disease control in this region.
Original languageEnglish
JournalJournal of Virological Methods
Volume237
Pages (from-to)114-120
Number of pages7
ISSN0166-0934
DOIs
StatePublished - 2016
CitationsWeb of Science® Times Cited: 3

    Keywords

  • Virology, East Africa, Foot-and-mouth-disease virus, Real-time RT-PCR, Serotype-specific
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