EPPO Bulletin (2005) 35, 133-140

J.W. Roenhorst, C.C.C. Jansen, L.F.F. Kox, E.G. de Haan, G.W. van den Bovenkamp, N. Boonham, T. Fisher and R.A. Mumford (2005)
Application of real-time RT-PCR for large-scale testing of potato for Potato spindle tuber pospiviroid
EPPO Bulletin 35 (1), 133-140
Abstract: The real-time RT-PCR protocol of Boonham and coworkers performed extremely well in a recent ring test, comparing different methods for detection of Potato spindle tuber pospiviroid (PSTVd) in several laboratories. Since, in addition, real-time PCR technology has proved suitable for high-throughput testing, this method was chosen as the starting point for the development of a protocol for the large-scale testing of potato. The initial experiments focused on the specificity of the primers and probes with regard to different isolates of PSTVd and other (pospi-) viroids. Further experiments were performed with leaf material from both primarily and secondarily infected plants. The parameters studied were sampling position, growing-on temperature and bulking rate. In addition, different grinding, nucleic-acid extraction and disinfection methods were compared. To monitor false negatives and positives, different controls were included and tested in duplex and triplex formats. The final protocol was tested using a hundred samples from the Dutch potato-monitoring programme. The results of this pilot experiment were promising. Future plans include the development of a protocol for direct tuber testing and inter-laboratory ring testing of the protocols.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Linda F.F. Kox, Neil Boonham

Research topic(s) for pests/diseases/weeds:
identification/taxonomy
surveys/sampling/distribution

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Potato spindle tuber viroid	Potato (Solanum tuberosum)