Devolpment of a nested PCR protocol for detection of olive-infecting viruses in crude extracts

A reverse transcription nested polymerase chain reaction (RT-n-PCR) was developed and applied to the detection of Arabis mosaic virus (ArMV), Cherry leaf roll virus (CLRV), Olive latent virus-1 (OLV-1), Olive latent virus-2 (OLV-2), Olive latent ringspot virus (OLRSV) and Strawberry latent ringspot virus (SLRV) in naturally infected olive cuttings. The method was more sensitive and reliable than conventional RT-PCR. This increased sensitivity allowed virus detection in samples that were negative to the first round of RT-PCR amplification. Coupled with simple sample preparation from cortical scrapings of olive cuttings, this protocol should be useful in routine testing for olive viruses.