Detection: Candidatus phytoplasma vitis, responsible of Flavescence dorée and Candidatus phytoplasma solani, responsible of Bois noir on grapevine, with distinction of the two phytoplasma.
Matrix: Grapevine leaves, preferably primary veins and petioles. Other parts of grapevine tissues can be analyzed, for example vine stocks, or insects’ vectors, using specific nucleic acids extraction’s methods.
Other information: kit based on an analysis method developed by the company International Plant Analysis and Diagnostics (www.ipadlab.eu).
The primers and probes were designed on the rpl14 FD gene and the rpl22-rps3 BN gene. The primers and probes sequences and their use in diagnostic tests are the subject of a PCT patent application by IpadLab (PCT/IB2010/053563). This Triplex Real-Time PCR kit offers a specific and sensitive method to detect simultaneously 16SrV phytoplasmas including FD and 16SrXII phytoplasmas including BN, as well as an endogenous control designed on the COX gene.
Taq-Man® technology, FAM (FD), VIC (BN) and Cy5 (Internal Control) fluorophores.
Validation data of the method are available from a test performance study realized in 2013 (Euphresco Grafdepi project). This method is also referred in the Appendix 6 of the PM7/079 (2) Grapevine Flavescence dorée phytoplasma, European and Mediterranean Plant Protection Organization Bulletin (2016) 46 (1), 78-93.
Kit referred in PM7/079 (2).
Included reagents: Direct Master Mix, Negative Control, Positive Control.