Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses
Globally, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) occur frequently and in combination cause sweetpotato virus disease (SPVD). Many viral diseases are economically important and negatively impact the production and movement of germplasm across regions...
| Autores principales: | , , , , , , |
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| Formato: | Journal Article |
| Lenguaje: | Inglés |
| Publicado: |
Elsevier
2021
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| Materias: | |
| Acceso en línea: | https://hdl.handle.net/10568/115161 |
| _version_ | 1855531996180119552 |
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| author | Wanjala, Bramwel W. Ateka, E.M. Miano, D.W. Fuentes, S. Pérez, A. Low, Jan W. Kreuze, Jan F. |
| author_browse | Ateka, E.M. Fuentes, S. Kreuze, Jan F. Low, Jan W. Miano, D.W. Pérez, A. Wanjala, Bramwel W. |
| author_facet | Wanjala, Bramwel W. Ateka, E.M. Miano, D.W. Fuentes, S. Pérez, A. Low, Jan W. Kreuze, Jan F. |
| author_sort | Wanjala, Bramwel W. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Globally, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) occur frequently and in combination cause sweetpotato virus disease (SPVD). Many viral diseases are economically important and negatively impact the production and movement of germplasm across regions. Rapid detection of viruses is critical for effective control. Detection and quantification of viruses directly from sweetpotato remains a challenge. Current diagnostic tests are not sensitive enough to reliably detect viruses directly from the plant or require expensive laboratory equipment and expertise to perform. We developed a simple and rapid loop‐mediated isothermal amplification (LAMP) assay for the detection of SPFMV, SPCSV and begomoviruses related to sweet potato leaf curl virus (SPLCV). Laboratory validation recorded 100% diagnostic sensitivity for all three viruses. The LAMP assays were customized for field testing using a lyophilized thermostable isothermal master mix in a ready-to-use form that required no cold chain. The average time to positivity (TTP) was: SPFMV 5-30 min., SPCSV 15-43 mins and begomoviruses 28-45 mins. LAMP on-site testing results were comparable to PCR and RT-PCR confirmatory laboratory tests. The LAMP assay is a powerful tool for rapid sweetpotato virus detection at a reasonable cost and thus could serve as quality control systems for planting materials. |
| format | Journal Article |
| id | CGSpace115161 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | Elsevier |
| publisherStr | Elsevier |
| record_format | dspace |
| spelling | CGSpace1151612024-05-01T08:18:25Z Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses Wanjala, Bramwel W. Ateka, E.M. Miano, D.W. Fuentes, S. Pérez, A. Low, Jan W. Kreuze, Jan F. sweet potatoes detection viroses begomovirus Globally, Sweet potato feathery mottle virus (SPFMV) and Sweet potato chlorotic stunt virus (SPCSV) occur frequently and in combination cause sweetpotato virus disease (SPVD). Many viral diseases are economically important and negatively impact the production and movement of germplasm across regions. Rapid detection of viruses is critical for effective control. Detection and quantification of viruses directly from sweetpotato remains a challenge. Current diagnostic tests are not sensitive enough to reliably detect viruses directly from the plant or require expensive laboratory equipment and expertise to perform. We developed a simple and rapid loop‐mediated isothermal amplification (LAMP) assay for the detection of SPFMV, SPCSV and begomoviruses related to sweet potato leaf curl virus (SPLCV). Laboratory validation recorded 100% diagnostic sensitivity for all three viruses. The LAMP assays were customized for field testing using a lyophilized thermostable isothermal master mix in a ready-to-use form that required no cold chain. The average time to positivity (TTP) was: SPFMV 5-30 min., SPCSV 15-43 mins and begomoviruses 28-45 mins. LAMP on-site testing results were comparable to PCR and RT-PCR confirmatory laboratory tests. The LAMP assay is a powerful tool for rapid sweetpotato virus detection at a reasonable cost and thus could serve as quality control systems for planting materials. 2021-12 2021-09-26T03:14:32Z 2021-09-26T03:14:32Z Journal Article https://hdl.handle.net/10568/115161 en Open Access Elsevier Wanjala, B. W., Ateka, E. M., Miano, D. W., Fuentes, S., Perez, A., Low, J. W., & Kreuze, J. F. (2021). Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses. Journal of Virological Methods. ISSN 1879-0984, 114301. |
| spellingShingle | sweet potatoes detection viroses begomovirus Wanjala, Bramwel W. Ateka, E.M. Miano, D.W. Fuentes, S. Pérez, A. Low, Jan W. Kreuze, Jan F. Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title | Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title_full | Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title_fullStr | Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title_full_unstemmed | Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title_short | Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses |
| title_sort | loop mediated isothermal amplification assays for on site detection of the main sweetpotato infecting viruses |
| topic | sweet potatoes detection viroses begomovirus |
| url | https://hdl.handle.net/10568/115161 |
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