ISHS Acta Horticulturae 386: XVI International Symposium on Fruit Tree Virus diseases
DETECTION OF PLUM POX VIRUS USING A SIMPLIFIED POLYMERASE CHAIN REACTION PROCEDURE |
| Authors: | L.M. Corvo, M. Sousa Santos, G. Nolasco |
| Abstract:
After the occurence of Plum Pox Virus (PPV) in Portugal, in 1984, control measures based on surveys by visual inspection, ELISA assays and biological methods, were applied. The development of easy, rapid, reliable and standardized detection methods is one of the priorities whenever the control of quarantine pathogens is concerned. Thus, in this work, the immunocapture - reverse transcriptional polymerase chain reaction (IC/RT - PCR) molecular method has been carried out following the procedure suggested by Wetzel et al., (1992) and Nolasco et al., (1992), but with the reverse transcription of the RNA performed directly on the material retained by solid-phase adsorbed antibodies, avoiding any previous thermal or chemical disruption of the virus particles. This strategy, in which the release of viral DNA seems to be coupled with the reverse transcription activity, was successfully developed by Nolasco et al. (1993) for the detection of several virus and other subviral pathogens. The results can be compared with those of the ELISA assay. The high sensitivity of this technique together with its speed and simplicity (not more laborious than ELISA), allows the-improvement of PPV control, mainly in mother trees and in intercommunity exchanges of plant material. A first attempt to characterize PPV isolates using this method complemented by the restriction analysis of the amplified fragment with the endonuclease Rsa I is described. | |
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