ISHS Acta Horticulturae 235: XIV International Symposium on Fruit Tree Virus Diseases
USE OF IN VITRO TRANSCRIBED RNAS FOR PLUM POX VIRUS DETECTION IN A DOT BLOT HYBRIDIZATION ASSAY |
| Authors: | C. Varveri, T. Candresse, M. Ravelonandro, G. Macquaire, J. Dunez |
| Abstract:
We have developed a dot blot molecular hybridization assay for the detection of plum pox virus, the causal agent of the sharka disease of stone fruit trees. The 800 nucleotide long probe used in this study was derived from the 3' region of the viral RNA and covered about 65% of the coat protein gene coding sequence. Use of 32P labeled, in vitro transcribed RNA probes as compared to 32P labeled, nick translated DNA probes greatly improves the sensitivity of the assay. The detection limit of the assay was 0.4 ng virus per milliliter, a value 10 times better than that obtained with ELISA. In a large scale comparative study, molecular hybridization proved to be much more sensitive than ELISA, detecting the virus in 99% of the ELISA positive samples and in 76% of the ELISA negative samples collected from infected trees. | |
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