Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples
The aim of this study was to characterise the performance of new molecular methods for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) and to provide validation data in comparison to the assays mentioned in official diagnostic protocols and being currently used. Eleven...
| Autores principales: | , , , , , , , , , , , , , , , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
2019
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| Acceso en línea: | http://hdl.handle.net/20.500.11939/6222 |
| _version_ | 1855032388548034560 |
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| author | Loreti, Stefania Cunty, Amandine Pucci, Nicoletta Chabirand, Aude Stefani, Emilio Abelleira, Adela Balestra, Giorgio M. Cornish, Deirdre A. Gaffuri, Francesca Giovanardi, Davide Gottsberger, Richard A. Holeva, Maria C. Karahan, Aynur Karafla, Charikleia D. Mazzaglia, Angelo Taylor, Robert Cruz, Leonor López, María M. Vanneste, Joel L. Poliakoff, Francoise |
| author_browse | Abelleira, Adela Balestra, Giorgio M. Chabirand, Aude Cornish, Deirdre A. Cruz, Leonor Cunty, Amandine Gaffuri, Francesca Giovanardi, Davide Gottsberger, Richard A. Holeva, Maria C. Karafla, Charikleia D. Karahan, Aynur Loreti, Stefania López, María M. Mazzaglia, Angelo Poliakoff, Francoise Pucci, Nicoletta Stefani, Emilio Taylor, Robert Vanneste, Joel L. |
| author_facet | Loreti, Stefania Cunty, Amandine Pucci, Nicoletta Chabirand, Aude Stefani, Emilio Abelleira, Adela Balestra, Giorgio M. Cornish, Deirdre A. Gaffuri, Francesca Giovanardi, Davide Gottsberger, Richard A. Holeva, Maria C. Karahan, Aynur Karafla, Charikleia D. Mazzaglia, Angelo Taylor, Robert Cruz, Leonor López, María M. Vanneste, Joel L. Poliakoff, Francoise |
| author_sort | Loreti, Stefania |
| collection | ReDivia |
| description | The aim of this study was to characterise the performance of new molecular methods for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) and to provide validation data in comparison to the assays mentioned in official diagnostic protocols and being currently used. Eleven molecular tests for the Psa detection were compared in an inter-laboratory comparison where each laboratory had to analyse the same panel of samples consisting of thirteen Psa-spiked kiwifruit wood extracts. Laboratories had to perform also isolation from the wood extracts. Data from this interlaboratory test performance study (TPS) was statistically analysed to assess the performance of each method. In order to provide complete validation data, both for detection and identification, this TPS was supplemented by a further study of identification from pure culture of phylogenetically closely related Pseudomonas spp., Psa, and bacterial strains associated with kiwifruit. The results of both these studies showed that simplex-PCRs gave good results, whereas duplex-PCR and real-time PCR were the most reliable tools for detection and identification of Psa. Nested and multiplex-PCR gave false-positive results. The use of the most reliable detection test is suggested for routine analyses, but when Psa-free status needs to be accurately assessed, it is recommended that at least two detection tests are used. This work provides a wide comparison of the available diagnostic methods, giving new information for a possible revision of the official diagnostic protocols (e.g. European and Mediterranean Plant Protection Organization (EPPO) protocol PM7/120 for the detection of Psa). |
| format | article |
| id | ReDivia6222 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2019 |
| publishDateRange | 2019 |
| publishDateSort | 2019 |
| record_format | dspace |
| spelling | ReDivia62222025-04-25T14:46:33Z Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples Loreti, Stefania Cunty, Amandine Pucci, Nicoletta Chabirand, Aude Stefani, Emilio Abelleira, Adela Balestra, Giorgio M. Cornish, Deirdre A. Gaffuri, Francesca Giovanardi, Davide Gottsberger, Richard A. Holeva, Maria C. Karahan, Aynur Karafla, Charikleia D. Mazzaglia, Angelo Taylor, Robert Cruz, Leonor López, María M. Vanneste, Joel L. Poliakoff, Francoise The aim of this study was to characterise the performance of new molecular methods for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) and to provide validation data in comparison to the assays mentioned in official diagnostic protocols and being currently used. Eleven molecular tests for the Psa detection were compared in an inter-laboratory comparison where each laboratory had to analyse the same panel of samples consisting of thirteen Psa-spiked kiwifruit wood extracts. Laboratories had to perform also isolation from the wood extracts. Data from this interlaboratory test performance study (TPS) was statistically analysed to assess the performance of each method. In order to provide complete validation data, both for detection and identification, this TPS was supplemented by a further study of identification from pure culture of phylogenetically closely related Pseudomonas spp., Psa, and bacterial strains associated with kiwifruit. The results of both these studies showed that simplex-PCRs gave good results, whereas duplex-PCR and real-time PCR were the most reliable tools for detection and identification of Psa. Nested and multiplex-PCR gave false-positive results. The use of the most reliable detection test is suggested for routine analyses, but when Psa-free status needs to be accurately assessed, it is recommended that at least two detection tests are used. This work provides a wide comparison of the available diagnostic methods, giving new information for a possible revision of the official diagnostic protocols (e.g. European and Mediterranean Plant Protection Organization (EPPO) protocol PM7/120 for the detection of Psa). 2019-05-15T10:37:42Z 2019-05-15T10:37:42Z 2018 article Loreti, S.; Cunty, A.; Pucci, N.; Chabirand, A.; Stefani, E.; Abelleira, A.; Balestra, G. M.; Cornish, D. A.; Gaffuri, F.; Giovanardi, D.; Gottsberger, R. A.; Holeva, M.; Karahan, A.; Karafla, C. D.; Mazzaglia, A.; Taylor, R.; Cruz, L.; López, M. M.; Vanneste, J. L.; Poliakoff, F. (2018). Performance of diagnostic tests for the detection and identification of pseudomonas syringae pv. actinidiae (psa) from woody samples. European Journal of Plant Pathology, 152(3), 657-676. http://hdl.handle.net/20.500.11939/6222 10.1007/s10658-018-1509-5 en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ electronico |
| spellingShingle | Loreti, Stefania Cunty, Amandine Pucci, Nicoletta Chabirand, Aude Stefani, Emilio Abelleira, Adela Balestra, Giorgio M. Cornish, Deirdre A. Gaffuri, Francesca Giovanardi, Davide Gottsberger, Richard A. Holeva, Maria C. Karahan, Aynur Karafla, Charikleia D. Mazzaglia, Angelo Taylor, Robert Cruz, Leonor López, María M. Vanneste, Joel L. Poliakoff, Francoise Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title | Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title_full | Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title_fullStr | Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title_full_unstemmed | Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title_short | Performance of diagnostic tests for the detection and identification of Pseudomonas syringae pv. actinidiae (Psa) from woody samples |
| title_sort | performance of diagnostic tests for the detection and identification of pseudomonas syringae pv actinidiae psa from woody samples |
| url | http://hdl.handle.net/20.500.11939/6222 |
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