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    Multiplex RTPCR assays for the simultaneous detection of both RNA and DNA viruses infecting cassava and the common occurrence of mixed infections by two cassava brown streak viruses in East Africa

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    Authors
    Abarshi, M.M.
    Mohammed, I.
    Jeremiah, S.C.
    Legg, J.P.
    Kumar, P.L.
    Hillocks, R.J.
    Maruthi, M.N.
    Date
    2012-01
    Language
    en
    Type
    Journal Article
    Review status
    Peer Review
    Accessibility
    Limited Access
    Metadata
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    Citation
    Abarshi, M.M., Mohammed, I., Jeremiah, S.C., Legg, J., Kumar, P.L., Hillocks, R.J. & Maruthi, M.N. (2012). Multiplex RT-PCR assays for the simultaneous detection of both RNA and DNA viruses infecting cassava and the common occurrence of mixed infections by two cassava brown streak viruses in East Africa. Journal of Virological Methods, 179(1), 176-184.
    Permanent link to cite or share this item: http://hdl.handle.net/10568/77415
    DOI: https://dx.doi.org/10.1016/j.jviromet.2011.10.020
    Abstract/Description
    Uniplex and multiplex reverse transcription-polymerase chain reaction (RT-PCR) protocols were developed for the detection of cassava brown streak viruses (CBSVs) in single and mixed infections with cassava mosaic begomoviruses (CMBs) in a tropical crop plant, cassava (Manihot esculenta). CMBs contain ssDNA as their genome (genus Begomovirus, family Geminiviridae) while CBSVs are made up of positive sense ssRNA (genus Ipomovirus, family Potyviridae), and they cause the economically important cassava mosaic and cassava brown streak diseases, respectively, in sub-Saharan Africa. Diagnostic methodologies have long been available for CMBs but they are limited for CBSVs especially in mixed infections. In this study, the two CBSVs, Cassava brown streak virus (CBSV) and Cassava brown streak Uganda virus (CBSUV) occurring singly or in mixed infection with CMBs, African cassava mosaic virus and East African cassava mosaic virus were detected in a single RT-PCR using both previously described and newly designed virus-specific primers. These protocols were highly efficient for detecting CBSVs compared to the existing methods and have great potential to minimize sample handling and contamination. As well as improving the diagnosis of cassava viruses, the development of multiplex RT-PCR protocols have revealed the common occurrence of mixed infections by CBSV and CBSUV in cassava fields of Tanzania and Kenya, which was contrary to the common belief until recently that these two viruses have existed separately. These protocols have implications for diagnosis and epidemiological studies on cassava virus diseases in Eastern Africa.
    CGIAR Affiliations
    Roots, Tubers and Bananas
    AGROVOC Keywords
    CASSAVA; RNA; DNA
    Subjects
    CASSAVA; PLANT DISEASES
    Countries
    UGANDA; KENYA; ZAMBIA
    Regions
    AFRICA; EAST AFRICA; SOUTHERN AFRICA
    Investors/sponsors
    Bill & Melinda Gates Foundation
    Collections
    • IITA Journal Articles [1807]
    • RTB Journal Articles [656]

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