The plant pathogens that cause significant economic damage to agricultural crops are subject to phytosanitary regulations aimed at protecting the sanitary and quality of propagative material and production. The availability of official diagnostic protocols, therefore, is necessary for the uniformity of results obtained in different laboratories and to give them a legal value, essential for the application of the phytosanitary measures. Moreover, the entire international community is moving in this direction in order to standardize the diagnosis. The aim of the Italian Project ARNADIA, funded by the Ministry of Agriculture Food and Forestry, was to produce diagnostic protocols officially recognized at a national level for the identification of major plant pathogens. One of the sub-tasks of the ARNADIA Project planned the production of reference and validated protocols for the diagnosis of ‘Candidatus Phytoplasma mali’ and ‘Candidatus Phytoplasma prunorum’. For this purpose a working group was established, including the main Italian scientific institutions which are involved in the study of both phytoplasmas Several diagnostic protocols and reagents were compared in five different laboratories, using the same target and no-target reference samples. Protocols of PCR and real time PCR were selected and developed to determine the performance characteristics for validation under the standard UNI CEI EN ISO/IEC 17025 and EPPO PM7/84 and PM7/98. The results showed that the accuracy and analytical sensitivity of PCR performed with specific primers and nested PCR are comparable, whereas real time PCR results were more reliable. A ringtest among seven Italian laboratories of Phytosanitary Regional Services is currently to evaluate the concordance parameter of all selected protocols.
Validation of diagnostic protocolts for the detection of ‘Candidatus Phytoplasma prunorum’
BERTACCINI, ASSUNTA;PALTRINIERI, SAMANTA;
2012
Abstract
The plant pathogens that cause significant economic damage to agricultural crops are subject to phytosanitary regulations aimed at protecting the sanitary and quality of propagative material and production. The availability of official diagnostic protocols, therefore, is necessary for the uniformity of results obtained in different laboratories and to give them a legal value, essential for the application of the phytosanitary measures. Moreover, the entire international community is moving in this direction in order to standardize the diagnosis. The aim of the Italian Project ARNADIA, funded by the Ministry of Agriculture Food and Forestry, was to produce diagnostic protocols officially recognized at a national level for the identification of major plant pathogens. One of the sub-tasks of the ARNADIA Project planned the production of reference and validated protocols for the diagnosis of ‘Candidatus Phytoplasma mali’ and ‘Candidatus Phytoplasma prunorum’. For this purpose a working group was established, including the main Italian scientific institutions which are involved in the study of both phytoplasmas Several diagnostic protocols and reagents were compared in five different laboratories, using the same target and no-target reference samples. Protocols of PCR and real time PCR were selected and developed to determine the performance characteristics for validation under the standard UNI CEI EN ISO/IEC 17025 and EPPO PM7/84 and PM7/98. The results showed that the accuracy and analytical sensitivity of PCR performed with specific primers and nested PCR are comparable, whereas real time PCR results were more reliable. A ringtest among seven Italian laboratories of Phytosanitary Regional Services is currently to evaluate the concordance parameter of all selected protocols.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
