Progetto ARNADIA: definizione di protocolli nazionali di diagnosi per ‘Candidatus Phytoplasma mali’ e ‘Candidatus Phytoplasma prunorum’

The importance of diagnostic procedures harmonization, the contribution to improved transparency during the detection of regulated pests and the need of the resolution of disputes among trading partners suggested, in the frame of the Italian Project ARNADIA financed by the Ministry of Agriculture, to set up validated diagnostic protocols, officially approved and published at national level. To select a reference protocol for ‘Candidatus Phytoplasma mali’ and ‘Candidatus Phytoplasma prunorum’ detection different diagnostic methods, protocols and reagents have been compared in five laboratories, using the same target and non-target reference samples. Methods of conventional and real time PCR have been selected and developed to determine the performance characteristics for the validation under the standard ISO 17025. The results show that the accuracy and sensitivity of conventional and real time PCR are comparable, whereas real time RT-PCR is recommendable to increase the analytical sensitivity. The reference protocols are available under www.strateco.it.