Simultaneous detection of groundnut rosette assistor virus (GRAV), groundnut rosette virus (GRV) and satellite RNA (satRNA) in groundnuts using multiplex RT-PCR
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 %.
KeywordsGroundnut rosette disease Two-step multiplex RT-PCR Two-step RT-PCR
- 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
- 4.Murant AF (1989) Groundnut rosette assistor virus. CMI\AAB Descr Plant Viruses No 345Google Scholar
- 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
- 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
- 18.Sokal RR, Rohlf FJ (1995) Biometry: the principles and practice of statistics in biological research, 3rd edn. Freeman, New YorkGoogle Scholar