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    Rapid and specific detection of Yam mosaic virus by reverse-transcription recombinase polymerase amplification

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    S15ArtSilvaRapidInthomNodev.pdf (850.0Kb)
    Date
    2015
    Author
    Silva, G.
    Bömer, M.
    Nkere, C.
    Kumar, P.L.
    Seal, S.E.
    Type
    Journal Article
    Metadata
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    Abstract/Description
    Yam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratoriesYam mosaic virus (YMV; genus Potyvirus ) is considered to cause the most economically important viral disease of yams ( Dioscorea spp.) in West Africa which is the dominant region for yam production globally. Yams are a vegetatively propagated crop and the use of virus-free planting material forms an essential component of disease control. Current serological and PCR-based diagnostic methods for YMV are time consuming involving a succession of target detection steps. In this study, a novel assay for specific YMV detection is described that is based on isothermal reverse transcription-recombinase polymerase ampli- fication (RT-exoRPA). This test has been shown to be reproducible and able to detect as little as 14 pg/ l of purified RNA obtained from an YMV-infected plant, a sensitivity equivalent to that obtained with the reverse transcription-polymerase chain reaction (RT-PCR) in current general use. The RT-exoRPA assay has, however, several advantages over the RT-PCR; positive samples can be detected in less than 30 min, and amplification only requires a single incubation temperature (optimum 37 ? C). These features make the RT-exoRPA assay a promising candidate for adapting into a field test format to be used by yam breeding programmes or certification laboratories
    https://dx.doi.org/10.1016/j.jviromet.2015.06.011
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/880
    Digital Object Identifier (DOI)
    https://dx.doi.org/10.1016/j.jviromet.2015.06.011
    IITA Subjects
    Yam; Plant Diseases
    Agrovoc Terms
    Yams; Dioscorea; Mosaic Virus; Diagnosis
    Regions
    Africa; West Africa
    Countries
    Nigeria
    Journals
    Journal of Virological Methods
    Collections
    • Journal and Journal Articles4836
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