Articles
DETECTION OF PRUNE DWARF VIRUS IN SWEET CHERRY IN ISRAEL
Article number
472_27
Pages
249 – 256
Language
Abstract
Prune dwarf ilarvirus (PDV) was identified for the first time in Israel in several imported sweet cherry cultivars and one peach cultivar.
Double-antibody sandwich (DAS) and triple antibody sandwich (TAS) ELISA tests were successfully used for PDV detection during spring in most infected cultivars with one exception (cv.
Hedelfingen). The sensitivity and reliability of TAS-ELISA was higher than the polyclonal-based assay.
ELISA results correlated with graft-inoculation onto Shirofugen indicator plants.
Reverse-transcription (RT) and immunocapture (IC) polymerase chain reaction (PCR) amplification assays applied to PDV-infected sweet cherry cultivars yielded consistently a specific product (previously reported by Parakh et al., 1995) except for Hedelfingen.
Using diluted sap in IC-PCR resulted in a larger amount of amplified product and improved detection.
PCR assays were superior to ELISA, season-independent and generally allowed overcoming the uneven distribution of PDV in woody hosts, typical of ilarviruses.
In conclusion, we suggest to include, in addition to biological indexing, a two-step assay for detection of PDV in post-entry tests and certification schemes: i) ELISA, preferably a MAB-based assay, especially if large-scale testing is required; and ii) PCR for ELISA-negative and inclusive samples.
Double-antibody sandwich (DAS) and triple antibody sandwich (TAS) ELISA tests were successfully used for PDV detection during spring in most infected cultivars with one exception (cv.
Hedelfingen). The sensitivity and reliability of TAS-ELISA was higher than the polyclonal-based assay.
ELISA results correlated with graft-inoculation onto Shirofugen indicator plants.
Reverse-transcription (RT) and immunocapture (IC) polymerase chain reaction (PCR) amplification assays applied to PDV-infected sweet cherry cultivars yielded consistently a specific product (previously reported by Parakh et al., 1995) except for Hedelfingen.
Using diluted sap in IC-PCR resulted in a larger amount of amplified product and improved detection.
PCR assays were superior to ELISA, season-independent and generally allowed overcoming the uneven distribution of PDV in woody hosts, typical of ilarviruses.
In conclusion, we suggest to include, in addition to biological indexing, a two-step assay for detection of PDV in post-entry tests and certification schemes: i) ELISA, preferably a MAB-based assay, especially if large-scale testing is required; and ii) PCR for ELISA-negative and inclusive samples.
Authors
S. Spiegel, A. Rosner, Y. Tam, S. Zilkah, E. Faingersh, A. Rotbaum, L. Krizbai
Keywords
Ilarviruses, stone-fruits, sweet cherries, virus detection
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