Archivio Journal of Plant Pathology

Abstract:

Detection of Potato virus Y (PVY), one of the important potato viruses, was undertaken using various PCRbased techniques, i.e immunocapture reverse transcription polymerase chain reaction (IC-RT-PCR), direct binding RT-PCR (DB-RT-PCR) and print capture RTPCR (PC-RT-PCR). These techniques were evaluated for detection of PVY virions immobilized on solid supports. The 5’UTR and P1 region (969 bp) of the PVY genome were amplified using these techniques, which were compared with DAS-ELISA and nucleic acid spot hybridization (NASH) using fluorescein labeled cDNA probes. The detection limit of purified PVY in serially diluted healthy potato leaf sap was 10 ng for DAS-ELISA, 1 ng for NASH and DB-RT-PCR, 1pg for PC-RT-PCR, ICRT- PCR and standard RT-PCR. Of all techniques, PCRT- PCR proved to be the simplest, economical and reliable. It does not require antibodies for the capture of virions nor lengthy RNA extraction processes.