Development of a Quick Quantitative Real-Time PCR for the In Vivo Detection and Quantification of Peach latent mosaic viroid

Viroids are plant pathogens infecting a broad range of herbaceous and
tree crops. Among them, the Peach latent mosaic viroid (PLMVd)
infects mainly peach trees, causing a loss of production with no
curative options. Detecting this viroid is thus important for certification
procedures aiming to avoid the release of infected material into
orchards. Presented here is a complete detection method based on
reverse transcription (RT) followed by a quantitative real-time
polymerase chain reaction (PCR). New primers were selected and
optimal reaction conditions determined for routine application of the
method. The technique is 105 times more sensitive than the endpoint
RT-PCR used for PLMVd detection, and permits earlier detection of
PLMVd in infected plants. The quick, low-cost extraction procedure
used and the quality of the results obtained make this method suitable
for routine testing.