Validation of a microarrays protocol for detection and genotyping isolates of Plum Pox
Pasquini, G.; Faggioli, F.; Luigi, M.: Gentili, A.; Hadidi, A.; Canini, I.; Gabriele, L.; Czosnek, H.; Tiberini, A.; Ça?layan, K.; Mazyad, H.; Anfoka, G.; Barba, M.
Proceedings of 21st International Conference on Virus and other Graft Transmissible Diseases of Fruit Crops
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julius Kuhn Archiv
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|Abstract in English:|
A genomic strategy for PPV identification has been recently developed (Pasquini et al., 2008). The method is based on
using a 70-mer oligonucleotide DNA microarray chip capable of simultaneously detecting and genotyping PPV strains.
Universal and specific probes have been identified and used with a sensitive protocol of hybridization using an indirect
fluorescent labelling of cDNA product with cyanine able to enhance the sensitivity of the virus detection avoiding the
use of the PCR amplification step. In order to evaluate the protocol fitness for diagnostic use, about 30 samples
belonging to a PPV isolates collection, including M, D, EA and C strains, have been used for its validation, that was
determined, estimating the performance criteria that include the following parameters: diagnostic sensitivity (D-SN),
diagnostic specificity (D-SP) and diagnostic accuracy (D-AC).