Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants
Erwinia amylovora, the fire blight pathogen, can survive in plants and produce latent infections that can lead to false negative results in routine diagnostic procedures, since the pathogen can be present at very low numbers under the detection limit of available techniques. A protocol combining dif...
| Main Authors: | , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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Springer
2020
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| Online Access: | http://hdl.handle.net/20.500.11939/6698 http://www.sipav.org/main/jpp/index.php/jpp/article/view/3912 |
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| author | Catalá-Senent, J. F. Penyalver, Javier Navarro, Luis Baranauskaite, L. Gorris, María T. López, María M. Marco-Noales, Ester |
| author_browse | Baranauskaite, L. Catalá-Senent, J. F. Gorris, María T. López, María M. Marco-Noales, Ester Navarro, Luis Penyalver, Javier |
| author_facet | Catalá-Senent, J. F. Penyalver, Javier Navarro, Luis Baranauskaite, L. Gorris, María T. López, María M. Marco-Noales, Ester |
| author_sort | Catalá-Senent, J. F. |
| collection | ReDivia |
| description | Erwinia amylovora, the fire blight pathogen, can survive in plants and produce latent infections that can lead to false negative results in routine diagnostic procedures, since the pathogen can be present at very low numbers under the detection limit of available techniques. A protocol combining different techniques included in the EPPO diagnostic standard for E. amylovora (2013) was designed in order to determine the minimum amount of plants necessary for detecting the pathogen in asymptomatic plant material. To this aim, single samples of flowers and shoots from pear, apple, and loquat were spiked with an E. amylovora strain at different concentrations, and then analyzed by isolation, real-time PCR and ELISA-DASI. Thereafter, using the single spiked samples that were positive for any of the techniques used, composite spiked samples were prepared by adding to single spiked samples increasing amounts of plant material. Interestingly, whole results show that the most robust technique, less affected by the amount of plant material, was the real-time PCR after enrichment, since 101 -102 cfu ml-1 of E. amylovora could be detected in most cases, both in flowers and in shoots, regardless of the amount of simulated plants analyzed, 3, 5, 10 or 20. The enrichment was confirmed as a necessary step in the analysis of bulked samples to achieve a greater sensitivity. This study is a relevant contribution for programs of extensive screening surveys for prevention of fire blight disease, since it shows that samples can be processed as a pool, improving the detection of E. amylovora in asymptomatic samples. |
| format | Artículo |
| id | ReDivia6698 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2020 |
| publishDateRange | 2020 |
| publishDateSort | 2020 |
| publisher | Springer |
| publisherStr | Springer |
| record_format | dspace |
| spelling | ReDivia66982025-04-25T14:47:41Z Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants Catalá-Senent, J. F. Penyalver, Javier Navarro, Luis Baranauskaite, L. Gorris, María T. López, María M. Marco-Noales, Ester EPPO ELISA-DASI Enrichment H20 Plant diseases Fire blight Latent infections Surveys Real-time PCR Erwinia amylovora, the fire blight pathogen, can survive in plants and produce latent infections that can lead to false negative results in routine diagnostic procedures, since the pathogen can be present at very low numbers under the detection limit of available techniques. A protocol combining different techniques included in the EPPO diagnostic standard for E. amylovora (2013) was designed in order to determine the minimum amount of plants necessary for detecting the pathogen in asymptomatic plant material. To this aim, single samples of flowers and shoots from pear, apple, and loquat were spiked with an E. amylovora strain at different concentrations, and then analyzed by isolation, real-time PCR and ELISA-DASI. Thereafter, using the single spiked samples that were positive for any of the techniques used, composite spiked samples were prepared by adding to single spiked samples increasing amounts of plant material. Interestingly, whole results show that the most robust technique, less affected by the amount of plant material, was the real-time PCR after enrichment, since 101 -102 cfu ml-1 of E. amylovora could be detected in most cases, both in flowers and in shoots, regardless of the amount of simulated plants analyzed, 3, 5, 10 or 20. The enrichment was confirmed as a necessary step in the analysis of bulked samples to achieve a greater sensitivity. This study is a relevant contribution for programs of extensive screening surveys for prevention of fire blight disease, since it shows that samples can be processed as a pool, improving the detection of E. amylovora in asymptomatic samples. 2020-10-27T08:15:57Z 2020-10-27T08:15:57Z 2017 article publishedVersion Català-Senent, J. F., Peñalver, J., Navarro, I., Baranauskaite, L., Gorris, M. T., López, M. M., & Marco-Noales, E. (2017). Laboratory-scale evaluation of composite spiked samples for Erwinia Amylovora detection in asymptomatic plants. Journal of Plant Pathology, 99, 105-110. 1125-4653 (Print ISSN) 2239-7264 (e-ISSN) http://hdl.handle.net/20.500.11939/6698 10.4454/jpp.v99i0.3912 http://www.sipav.org/main/jpp/index.php/jpp/article/view/3912 en Atribución-NoComercial-SinDerivadas 3.0 España http://creativecommons.org/licenses/by-nc-nd/3.0/es/ openAccess Springer electronico |
| spellingShingle | EPPO ELISA-DASI Enrichment H20 Plant diseases Fire blight Latent infections Surveys Real-time PCR Catalá-Senent, J. F. Penyalver, Javier Navarro, Luis Baranauskaite, L. Gorris, María T. López, María M. Marco-Noales, Ester Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title | Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title_full | Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title_fullStr | Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title_full_unstemmed | Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title_short | Laboratory-scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| title_sort | laboratory scale evaluation of composite spiked samples for erwinia amylovora detection in asymptomatic plants |
| topic | EPPO ELISA-DASI Enrichment H20 Plant diseases Fire blight Latent infections Surveys Real-time PCR |
| url | http://hdl.handle.net/20.500.11939/6698 http://www.sipav.org/main/jpp/index.php/jpp/article/view/3912 |
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