Archives of Virology

, Volume 159, Issue 11, pp 3059–3062 | Cite as

Simultaneous detection of groundnut rosette assistor virus (GRAV), groundnut rosette virus (GRV) and satellite RNA (satRNA) in groundnuts using multiplex RT-PCR

  • S. Anitha
  • E. S. Monyo
  • P. Okori
Brief Report


Groundnut rosette disease (GRD) is the most devastating disease of groundnuts in sub-Saharan Africa. The disease is caused by synergistic interactions between viruses and virus-like pathogens: groundnut rosette assistor virus (GRAV), groundnut rosette virus (GRV) and a satellite RNA (satRNA). The multi-pathogenic nature of GRD requires efficient diagnostic systems for plant breeding and pathology work. Currently, TAS-ELISA and RT-PCR are used to detect all three pathogens. This approach is time-consuming, expensive and not easily amenable to high throughput. A multiplex PCR-based approach was developed to detect all three pathogens at once, reducing diagnostics costs and time by two thirds. The technique is highly robust and amenable to high throughput, with sensitivity and specificity values of 88 % and 100 %, respectively. The positive predictive value for the technique is 100 %, and the negative predictive value is 90.6 %.


Groundnut rosette disease Two-step multiplex RT-PCR Two-step RT-PCR 



This study was funded through the generous support of the McKnight Foundation and the CGIAR Research Programme on Grain Legumes.


  1. 1.
    Waliyar F, Kumar PL, Ntare BR, Monyo E, Nigam SN, Reddy AS, Osiru M, Diallo AT (2007) Groundnut rosette disease and its management. Information Bulletin 75. International Crops Research Institute for the Semi-Arid Tropics, Patancheru, India, p 36Google Scholar
  2. 2.
    Casper R, Meyer SM, Lesemann DE, Reddy DVR, Rajeswari R, Misari SM, Subbarayudu SS (1983) Detection of a luteovirus in groundnut rosette diseased groundnuts (Arachis hypogaea) by enzyme-linked immunosorbent assay and immunoelectron microscopy. Phytopathol Z 108:12–17CrossRefGoogle Scholar
  3. 3.
    Reddy DVR, Murant AF, Duncan GH, Ansa OA, Demski JW, Kuhn CW (1985) Viruses associated with chlorotic rosette and green rosette diseases of groundnut in Nigeria. Ann Appl Biol 107:57–64CrossRefGoogle Scholar
  4. 4.
    Murant AF (1989) Groundnut rosette assistor virus. CMI\AAB Descr Plant Viruses No 345Google Scholar
  5. 5.
    Storey HH, Bottomley BA (1928) The rosette disease of peanuts (Arachis hypogaea L.). Ann Appl Biol 5:26–45CrossRefGoogle Scholar
  6. 6.
    Storey HH, Ryland AK (1955) Transmission of groundnut rosette virus. Ann App Biol 43:423–432CrossRefGoogle Scholar
  7. 7.
    Storey HH, Ryland AK (1957) Viruses causing Rosette and other diseases in groundnuts. Ann App Biol 45:319–326CrossRefGoogle Scholar
  8. 8.
    Hull R, Adams AN (1968) Groundnut rosette and its assistor virus. Ann Appl Biol 62:139–145CrossRefGoogle Scholar
  9. 9.
    Rajeswari R, Murant AF, Massalski PR (1987) Use of monoclonal antibody to potato leaf roll virus for detecting groundnut rosette assistor virus by ELISA. Ann Appl Biol 111:353–358CrossRefGoogle Scholar
  10. 10.
    Naidu RA, Bottenburg H, Subrahmanyam P, Kimmins FM, Robinson DJ, Thresh JM (1998) Epidemiology of groundnut rosette virus disease: current status and future research needs. Ann Appl Biol 132:525–548CrossRefGoogle Scholar
  11. 11.
    Blok VC, Ziegler A, Scott K, Dangora DB, Robinson DJ, Murant AF (1995) Detection of groundnut rosette umbravirus infections with radioactive and non-radioactive probes to its satellite RNA. Ann Appl Biol 117:379–384Google Scholar
  12. 12.
    Elnifro EM, Ashshi AM, Cooper RJ, Klapper PE (2000) Multiplex PCR: optimization and application in diagnostic virology. Clin Microbiol Rev 13:559–570PubMedCrossRefPubMedCentralGoogle Scholar
  13. 13.
    Bertolini E, Olmos A, Martinez MC, Gorris MT, Cambra M (2001) Single-step multiplex RT-PCR for simultaneous and colorimetric detection of six RNA viruses in olive trees. J Virol Methods 96:33–41PubMedCrossRefGoogle Scholar
  14. 14.
    Hauser S, Weber C, Vetter G, Stevens M, Beuve M, Lemaire O (2000) Improved detection and differentiation of poleroviruses infecting beet or rape by multiplex RT-PCR. J Virol Methods 89:11–21PubMedCrossRefGoogle Scholar
  15. 15.
    Olorunju RE, Kuhn CW, Demski JW, Misari SM, Ansa OA (1991) Resistance in groundnut to mixed infections of groundnut rosette virus (GRV) and groundnut rosette assistor virus (GRAV), and to infection by GRV alone. In: Reddy DVR, McDonald D (eds) Groundnut rosette disease in Africa, ICRISAT, India, pp 18–20Google Scholar
  16. 16.
    Naidu RA, Robinson DJ, Kimmins FM (1998) Detection of each of the casual agents of groundnut rosette disease in plants and vector aphids by RT-PCR. J Virol Methods 76:9–18PubMedCrossRefGoogle Scholar
  17. 17.
    Kulkarni S, Singh P, Memon A, Nataraj G, Kanade S, Kelkar R, Rajan MGR (2012) An in-house multiplex PCR test for the detection of mycobacterium tuberculosis, its validation and comparison with a single target TB-PCR kit. Indian J Med Res 135:788–794PubMedPubMedCentralGoogle Scholar
  18. 18.
    Sokal RR, Rohlf FJ (1995) Biometry: the principles and practice of statistics in biological research, 3rd edn. Freeman, New YorkGoogle Scholar
  19. 19.
    Murant AF, Rajeswari R, Robinson DJ, Raschke JH (1988) A satellite RNA of groundnut rosette virus that is largely responsible for symptoms of groundnut rosette disease. J Gen Virol 69:1479–1486CrossRefGoogle Scholar
  20. 20.
    Murant AF, Kumar IK (1990) Different variants of the satellite RNA of groundnut rosette virus are responsible for the chlorotic and green forms of groundnut rosette disease. Ann Appl Biol 117:85–92CrossRefGoogle Scholar
  21. 21.
    Taliansky ME, Robinson DJ, Murrant AF (1996) Complete nucleotide sequences and organization of the RNA sequence of groundnut rosette umbra virus. J Gen Virol 77:2335–2345PubMedCrossRefGoogle Scholar
  22. 22.
    Scott KP, Farmer MT, Robinson DJ, Torrance L, Murrant AF (1996) Comparison of the coat protein of groundnut rosette assistor virus with those of other luteo virus. Ann Appl Biol 128:77–83CrossRefGoogle Scholar
  23. 23.
    Blok VL, Ziegler A, Robinson DJ, Murrant AF (1994) Sequences of 10 variants of the satellite like RNA-3 of groundnutrosette virus. Virol 202:25–32CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Wien 2014

Authors and Affiliations

  1. 1.International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)LilongweMalawi
  2. 2.International Crops Research Institute for the Semi-Arid Tropics (ICRISAT)NairobiKenya

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