A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1
Grapevine (Vitis vinifera L.) is one of the most important crops in the world due to its economic and social impact. Like many other crops, grapevine is susceptible to different types of diseases caused by pathogenic microorganisms. Grapevine leafroll-associated virus 1 (GLRaV-1) is a virus associat...
| Main Authors: | , , , , , , |
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| Format: | Artículo |
| Language: | Inglés |
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MDPI
2023
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| Online Access: | https://hdl.handle.net/20.500.11939/8538 https://www.mdpi.com/2223-7747/12/4/876 |
| _version_ | 1855492509590880256 |
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| author | Morán, Félix Olmos, Antonio Glasa, Miroslav Bueno Da Silva, Marilia Maliogka, Varvara Wetzel, Thierry Ruiz-García, Ana B. |
| author_browse | Bueno Da Silva, Marilia Glasa, Miroslav Maliogka, Varvara Morán, Félix Olmos, Antonio Ruiz-García, Ana B. Wetzel, Thierry |
| author_facet | Morán, Félix Olmos, Antonio Glasa, Miroslav Bueno Da Silva, Marilia Maliogka, Varvara Wetzel, Thierry Ruiz-García, Ana B. |
| author_sort | Morán, Félix |
| collection | ReDivia |
| description | Grapevine (Vitis vinifera L.) is one of the most important crops in the world due to its economic and social impact. Like many other crops, grapevine is susceptible to different types of diseases caused by pathogenic microorganisms. Grapevine leafroll-associated virus 1 (GLRaV-1) is a virus associated with grapevine leafroll disease and it is considered at the national and European level as a pathogen that must be absent in propagative plant material. For this reason, the availability of specific, sensitive and reliable detection techniques to ascertain the sanitary status of the plants is of great importance. The objective of this research was the development of a new GLRaV-1 detection method based on a TaqMan quantitative real-time RT-PCR targeted to the coat protein genomic region and including a host internal control in a duplex reaction. To this end, three new GLRaV-1 full genomes were recovered by HTS and aligned with all sequences available in the databases. The method has been validated following EPPO standards and applied for the diagnosis of field plant material and transmission vectors. The new protocol designed has turned out to be highly sensitive as well as much more specific than the current available methods for the detection and absolute quantitation of GLRaV-1 viral titer. |
| format | Artículo |
| id | ReDivia8538 |
| institution | Instituto Valenciano de Investigaciones Agrarias (IVIA) |
| language | Inglés |
| publishDate | 2023 |
| publishDateRange | 2023 |
| publishDateSort | 2023 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | ReDivia85382025-04-25T14:49:03Z A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 Morán, Félix Olmos, Antonio Glasa, Miroslav Bueno Da Silva, Marilia Maliogka, Varvara Wetzel, Thierry Ruiz-García, Ana B. grapevine leafroll virus HTS diagnostics GLD PCR H20 Plant diseases Grapevine leaf roll virus Grapevine (Vitis vinifera L.) is one of the most important crops in the world due to its economic and social impact. Like many other crops, grapevine is susceptible to different types of diseases caused by pathogenic microorganisms. Grapevine leafroll-associated virus 1 (GLRaV-1) is a virus associated with grapevine leafroll disease and it is considered at the national and European level as a pathogen that must be absent in propagative plant material. For this reason, the availability of specific, sensitive and reliable detection techniques to ascertain the sanitary status of the plants is of great importance. The objective of this research was the development of a new GLRaV-1 detection method based on a TaqMan quantitative real-time RT-PCR targeted to the coat protein genomic region and including a host internal control in a duplex reaction. To this end, three new GLRaV-1 full genomes were recovered by HTS and aligned with all sequences available in the databases. The method has been validated following EPPO standards and applied for the diagnosis of field plant material and transmission vectors. The new protocol designed has turned out to be highly sensitive as well as much more specific than the current available methods for the detection and absolute quantitation of GLRaV-1 viral titer. 2023-02-27T09:54:29Z 2023-02-27T09:54:29Z 2023 article publishedVersion Morán, F., Olmos, A., Glasa, M., Silva, M. B. D., Maliogka, V., Wetzel, T., & Ruiz-García, A. B. (2023). A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll-Associated Virus 1. Plants, 12(4), 876. 2223-7747 https://hdl.handle.net/20.500.11939/8538 10.3390/plants12040876 https://www.mdpi.com/2223-7747/12/4/876 en This research was funded by the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement, No 734736. This publication only reflects the authors’ view. The Agency is not responsible for any use that may be made of the information it contains. M.G. was supported by the grant APVV-18-0005 from the Slovak Research and Development Agency and the project VEGA 2/0030/20 from the Scientific Grant Agency of the Ministry of Education, Science, Research and Sport of the Slovak Republic and the Slovak Academy of Sciences info:eu-repo/grantAgreement/EC/H2020/734736/EU//VirFree Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ openAccess MDPI electronico |
| spellingShingle | grapevine leafroll virus HTS diagnostics GLD PCR H20 Plant diseases Grapevine leaf roll virus Morán, Félix Olmos, Antonio Glasa, Miroslav Bueno Da Silva, Marilia Maliogka, Varvara Wetzel, Thierry Ruiz-García, Ana B. A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title | A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title_full | A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title_fullStr | A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title_full_unstemmed | A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title_short | A Novel and Highly Inclusive Quantitative Real-Time RT-PCR Method for the Broad and Efficient Detection of Grapevine Leafroll Associated Virus 1 |
| title_sort | novel and highly inclusive quantitative real time rt pcr method for the broad and efficient detection of grapevine leafroll associated virus 1 |
| topic | grapevine leafroll virus HTS diagnostics GLD PCR H20 Plant diseases Grapevine leaf roll virus |
| url | https://hdl.handle.net/20.500.11939/8538 https://www.mdpi.com/2223-7747/12/4/876 |
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