Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols
Peach latent mosaic viroid (PLMVd) is an important pathogen that causes disease in peaches. Control of this viroid remains problematic because most PLMVd variants are symptomless, and although there are many detection tests in use, the reliability of PCR-based methods is compromised by the complex,...
| Autores principales: | , , , , , , , , , , , , , |
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| Formato: | article |
| Lenguaje: | Inglés |
| Publicado: |
MDPI
2023
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/20.500.11939/8671 https://doi.org/10.3390/plants12091802 |
| _version_ | 1855032833013186560 |
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| author | Luigi, Marta Taglienti, Anna Corrado, Carla Libia Cardoni, Marco Botti, Simona Bissani, Rita Casati, Paola Passera, Alessandro Miotti, Niccolò De-Jonghe, Kris Everaert, Ellen Olmos, Antonio Ruiz-García, Ana B. Faggioli, Francesco |
| author_browse | Bissani, Rita Botti, Simona Cardoni, Marco Casati, Paola Corrado, Carla Libia De-Jonghe, Kris Everaert, Ellen Faggioli, Francesco Luigi, Marta Miotti, Niccolò Olmos, Antonio Passera, Alessandro Ruiz-García, Ana B. Taglienti, Anna |
| author_facet | Luigi, Marta Taglienti, Anna Corrado, Carla Libia Cardoni, Marco Botti, Simona Bissani, Rita Casati, Paola Passera, Alessandro Miotti, Niccolò De-Jonghe, Kris Everaert, Ellen Olmos, Antonio Ruiz-García, Ana B. Faggioli, Francesco |
| author_sort | Luigi, Marta |
| collection | ReDivia |
| description | Peach latent mosaic viroid (PLMVd) is an important pathogen that causes disease in peaches. Control of this viroid remains problematic because most PLMVd variants are symptomless, and although there are many detection tests in use, the reliability of PCR-based methods is compromised by the complex, branched secondary RNA structure of the viroid and its genetic diversity. In this study, a duplex RT-qPCR method was developed and validated against two previously published single RT-qPCRs, which were potentially able to detect all known PLMVd variants when used in tandem. In addition, in order to simplify the sample preparation, rapid-extraction protocols based on the use of crude sap or tissue printing were compared with commercially available RNA purification kits. The performance of the new procedure was evaluated in a test performance study involving five participant laboratories. The new method, in combination with rapid-sample-preparation approaches, was demonstrated to be feasible and reliable, with the advantage of detecting all different PLMVd isolates/variants assayed in a single reaction, reducing costs for routine diagnosis. |
| format | article |
| id | ReDivia8671 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | ReDivia86712025-04-25T14:49:15Z Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols Luigi, Marta Taglienti, Anna Corrado, Carla Libia Cardoni, Marco Botti, Simona Bissani, Rita Casati, Paola Passera, Alessandro Miotti, Niccolò De-Jonghe, Kris Everaert, Ellen Olmos, Antonio Ruiz-García, Ana B. Faggioli, Francesco Peach latent mosaic viroid Validated detection test TPS Rapid-extraction methods RT-PCR Low-cost diagnostic tests Nucleic-Acid-Extraction Protocols H20 Plant diseases U30 Research methods Peach latent mosaic viroid (PLMVd) is an important pathogen that causes disease in peaches. Control of this viroid remains problematic because most PLMVd variants are symptomless, and although there are many detection tests in use, the reliability of PCR-based methods is compromised by the complex, branched secondary RNA structure of the viroid and its genetic diversity. In this study, a duplex RT-qPCR method was developed and validated against two previously published single RT-qPCRs, which were potentially able to detect all known PLMVd variants when used in tandem. In addition, in order to simplify the sample preparation, rapid-extraction protocols based on the use of crude sap or tissue printing were compared with commercially available RNA purification kits. The performance of the new procedure was evaluated in a test performance study involving five participant laboratories. The new method, in combination with rapid-sample-preparation approaches, was demonstrated to be feasible and reliable, with the advantage of detecting all different PLMVd isolates/variants assayed in a single reaction, reducing costs for routine diagnosis. 2023-07-03T07:06:27Z 2023-07-03T07:06:27Z 2023 article publishedVersion Luigi, M., Taglienti, A., Corrado, C. L., Cardoni, M., Botti, S., Bissani, R. et al. (2023). Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols. Plants, 12(9), 1802. 2223-7747 https://hdl.handle.net/20.500.11939/8671 10.3390/plants12091802 https://doi.org/10.3390/plants12091802 en This research received no external funding Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ openAccess MDPI electronico |
| spellingShingle | Peach latent mosaic viroid Validated detection test TPS Rapid-extraction methods RT-PCR Low-cost diagnostic tests Nucleic-Acid-Extraction Protocols H20 Plant diseases U30 Research methods Luigi, Marta Taglienti, Anna Corrado, Carla Libia Cardoni, Marco Botti, Simona Bissani, Rita Casati, Paola Passera, Alessandro Miotti, Niccolò De-Jonghe, Kris Everaert, Ellen Olmos, Antonio Ruiz-García, Ana B. Faggioli, Francesco Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title | Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title_full | Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title_fullStr | Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title_full_unstemmed | Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title_short | Development and Validation of a Duplex RT-qPCR for Detection of Peach Latent Mosaic Viroid and Comparison of Different Nucleic-Acid-Extraction Protocols |
| title_sort | development and validation of a duplex rt qpcr for detection of peach latent mosaic viroid and comparison of different nucleic acid extraction protocols |
| topic | Peach latent mosaic viroid Validated detection test TPS Rapid-extraction methods RT-PCR Low-cost diagnostic tests Nucleic-Acid-Extraction Protocols H20 Plant diseases U30 Research methods |
| url | https://hdl.handle.net/20.500.11939/8671 https://doi.org/10.3390/plants12091802 |
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