Detection: Plum pox virus responsible of Sharka disease on Prunus.
Matrix: Plant leaves, flowers and buds.
Other information: kit based on Olmos et al., 2005. A verification was performed by Qualiplante (data not published) and the performance characteristics of the kit are the same as the original publication.
The assay was developed for the detection of the main types of PPV: PPV-M and PPV-D, without any distinction. To design appropriate primers and probes, the nucleotide sequence flanked by the universal primers P1 and P2 (Wetzel et al., 1991) was selected. The specific amplification of the target sequence is a 76 bp DNA fragment. This sensitive method was applied successfully to plant material and to individual PPV vector.
The Real-Time RT-PCR test of Olmos et al., 2005 is recommended by ANSES (www.anses.fr) – ANSES/LSV/MA 043 – Version 2 – Décembre 2018 and by Progetto Aron-Arnadia – Armonizzazione Protocollo diagnostico per Plum pox virus (PPV), Progetto Strateco, Anno 2012.
Taq-Man® technology, FAM fluorophore.
Included reagents: Direct Master Mix, RT-Enzyme, Negative control, Positive control.