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    Rapid detection of potyviruses from crude plant extracts

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    U18ArtSilvaRapidInthomNodev.pdf (433.8Kb)
    Date
    2018-04-01
    Author
    Silva, G.
    Oyekanmi, J.
    Nkere, C.K.
    Bömer, M.
    Kumar, P.L.
    Seal, S.E.
    Type
    Journal Article
    Target Audience
    Scientists
    Metadata
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    Abstract/Description
    Potyviruses (genus Potyvirus; family Potyviridae) are widely distributed and represent one of the most economically important genera of plant viruses. Therefore, their accurate detection is a key factor in developing efficient control strategies. However, this can sometimes be problematic particularly in plant species containing high amounts of polysaccharides and polyphenols such as yam (Dioscorea spp.). Here, we report the development of a reliable, rapid and cost-effective detection method for the two most important potyviruses infecting yam based on reverse transcription-recombinase polymerase amplification (RT-RPA). The developed method, named ‘Direct RT-RPA’, detects each target virus directly from plant leaf extracts prepared with a simple and inexpensive extraction method avoiding laborious extraction of high-quality RNA. Direct RT-RPA enables the detection of virus-positive samples in under 30 min at a single low operation temperature (37 °C) without the need for any expensive instrumentation. The Direct RT-RPA tests constitute robust, accurate, sensitive and quick methods for detection of potyviruses from recalcitrant plant species. The minimal sample preparation requirements and the possibility of storing RPA reagents without cold chain storage, allow Direct RT-RPA to be adopted in minimally equipped laboratories and with potential use in plant clinic laboratories and seed certification facilities worldwide.
    http://dx.doi.org/10.1016/j.ab.2018.01.019
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/2757
    Digital Object Identifier (DOI)
    http://dx.doi.org/10.1016/j.ab.2018.01.019
    IITA Subjects
    Pests Of Plants; Yam
    Agrovoc Terms
    Yams; Potyvirus; Plant Viruses; Food Security; Recombinase Polymerase Amplification; Isothermal Amplification; Rapid Diagnostics; Crude Plant Extracts
    Regions
    Africa; West Africa
    Countries
    Cote D'Ivoire; Nigeria
    Journals
    Analytical Biochemistry
    Collections
    • Journal and Journal Articles4835
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