Articles

DIRECT NESTED RT-PCR FOR THE DETECTION OF PLUM POX VIRUS

Article number
657_20
Pages
155 – 158
Language
English
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.

Publication
Authors
C. Varveri
Keywords
diagnosis, PPV, crude extracts
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