Standardization and implementation of the RT-PCR methodology for the diagnostic of cassava viruses

The in vitro collection of the genus Manihot of the Genetic Resource Program (GRP) of the International Center of Tropical Agriculture (CIAT) (Fig. 1) is currently represented by 6,643 materials; which have been registered in the Multilateral System of Access and Benefit sharing of the International Treaty on Plant Genetic Resources for Food and Agriculture. These materials must be free of quarantine diseases for distribution to users worldwide. The health certification of the cassava collection material that is done in the Germplasm Health Laboratory (LSG) of CIAT focuses in evaluating quarantine type viruses (Table1). According to the latest research, in addition to evaluating other viruses (CsXV, CsCMV, CVMV), it was necessary to implement and standardize reliable molecular methods for the detection of virus associated with different diseases that occur causing symptoms in the roots and the aerial part of the plant (Calvert et al. 2008, Carvajal et al. 2014). This study presents the standardization and implementation of molecular RT-PCR diagnostic methodology using random primers for cDNA synthesis and then specific and / or generic primers, allowing us to evaluate all quarantine virus reported from a single cDNA synthesis reaction. Given that the methodology is based on a total nucleic acid extraction, the same extraction can be used for the diagnosis of DNA viruses and microorganisms, if necessary.