Targeting highly conserved 3′-untranslated region of pecluviruses for sensitive broad-spectrum detection and quantitation by RT-PCR and assessment of phylogenetic relationships

    Dieryck, B and Delfosse, P and Reddy, A S and Bragard, C (2010) Targeting highly conserved 3′-untranslated region of pecluviruses for sensitive broad-spectrum detection and quantitation by RT-PCR and assessment of phylogenetic relationships. Journal of Virological Methods, 169 (2). pp. 385-390. ISSN 0166-0934

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    Abstract

    The 3′-end region of many virus isolates has been shown to possess conserved sequences in addition to the presence of numerous genomic and subgenomic RNAs. Utilizing these sequences, a broad-spectrum reverse transcription-polymerase chain reaction protocol has been developed to detect all the known Indian peanut clump virus and Peanut clump virus isolates, that cause peanut clump diseases in West Africa and India. The primers were targeted at the highly conserved 3′-untranslated regions of the PCV RNA-1 and RNA-2. The conservation was confirmed by sequencing these untranslated regions of RNA-1 for six isolates and RNA-2 for one isolate. The conserved structure of the RNA-1 and RNA-2 was observed and the importance of this region for the virus survival was confirmed. The primers were also designed for virus quantitation using a Taqman®-based real-time RT-PCR. The use of RT-PCR and real-time quantitative RT-PCR improved the sensitivity of PCV detection compared to ELISA. RT-PCR also led to the detection of IPCV and PCV on two new natural hosts: Oldenlandia aspera and Vigna subterranea. Real-time RT-PCR is considered to be an ideal tool for identifying resistant sources to both IPCV and PCV

    Item Type: Article
    Divisions: UNSPECIFIED
    CRP: UNSPECIFIED
    Uncontrolled Keywords: Peanut clump virus; Broad-spectrum detection; Virus quantitation; 3′-Untranslated region
    Subjects: Mandate crops > Groundnut
    Depositing User: Ms K Syamalamba
    Date Deposited: 18 Mar 2014 09:15
    Last Modified: 18 Mar 2014 09:15
    URI: http://oar.icrisat.org/id/eprint/7650
    Official URL: http://dx.doi.org/10.1016/j.jviromet.2010.08.010
    Projects: UNSPECIFIED
    Funders: UNSPECIFIED
    Acknowledgement: UNSPECIFIED
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