Detection of grapevine viruses by RT-PCR of double stranded RNA templates.
The non-availability of adequate immunological reagents has prevented the use modern diagnostic techniques for all the viruses that are considered in grapevine certification programs. A study was carried out to determine the feasibility of using double stranded RNA (ds-RNA) as a template for polymerase chain reaction (PCR) amplification and detection of 8 grapevine viruses (rupestris stem pitting associated virus, RSPaV; grapevine virus A, GVA; grapevine virus B, GVB; and grapevine leafroll-associated virus, GLRV), aiming at the development of an integrated system for viral diagnosis. All the viruses could be detected in variable amounts in nursery material. Rupestris stem pitting associated virus (RSPaV 1) and Grapevine virus B were frequently isolated. A significant number of samples previously assayed by ELISA were found positive for GLRV 3.