ISHS Acta Horticulturae 657: XIX International Symposium on Virus and Virus-like Diseases of Temperate Fruit Crops - Fruit Tree Diseases
DIRECT NESTED RT-PCR FOR THE DETECTION OF PLUM POX VIRUS |
| Author: | C. Varveri |
| Keywords: | diagnosis, PPV, crude extracts |
| Abstract:
The direct use of centrifuged plant extracts in water (1:20, w/v) was applied in a two-step RT-PCR procedure for the detection of Plum pox virus. The virus could be detected in apricot, peach and plum extracts with a sensitivity, however, that depending on the host was 10-1000 times inferior to that of the standard immunocapture (IC) procedure. In addition, non-specific reactions with the healthy control appeared in the case of plum extracts. These disadvantages were overcome by the use of nested RT-PCR in a single closed tube. Neither inhibition with healthy plant extracts nor non-specific reactions with the healthy controls were observed, clear-cut reactions were obtained and the sensitivity was at least similar to that of IC-RT-PCR. The independence from an immunocapture step and the extreme simplicity of sample preparation made this procedure most easy and quick to perform. | |
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