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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...

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Autores principales: Wanjala, Bramwel W., Ateka, Elijah M., Miano, Douglas W., Fuentes, Segundo, Perez, Ana, Low, Jan W., Kreuze, Jan F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier/North-Holland Biomedical Press 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8543070/
https://www.ncbi.nlm.nih.gov/pubmed/34560111
http://dx.doi.org/10.1016/j.jviromet.2021.114301
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author Wanjala, Bramwel W.
Ateka, Elijah M.
Miano, Douglas W.
Fuentes, Segundo
Perez, Ana
Low, Jan W.
Kreuze, Jan F.
author_facet Wanjala, Bramwel W.
Ateka, Elijah M.
Miano, Douglas W.
Fuentes, Segundo
Perez, Ana
Low, Jan W.
Kreuze, Jan F.
author_sort Wanjala, Bramwel W.
collection PubMed
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 the 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 min s 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.
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spelling pubmed-85430702021-12-01 Loop-Mediated Isothermal Amplification assays for on-site detection of the main sweetpotato infecting viruses Wanjala, Bramwel W. Ateka, Elijah M. Miano, Douglas W. Fuentes, Segundo Perez, Ana Low, Jan W. Kreuze, Jan F. J Virol Methods Article 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 the 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 min s 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. Elsevier/North-Holland Biomedical Press 2021-12 /pmc/articles/PMC8543070/ /pubmed/34560111 http://dx.doi.org/10.1016/j.jviromet.2021.114301 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Wanjala, Bramwel W.
Ateka, Elijah M.
Miano, Douglas W.
Fuentes, Segundo
Perez, Ana
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 Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8543070/
https://www.ncbi.nlm.nih.gov/pubmed/34560111
http://dx.doi.org/10.1016/j.jviromet.2021.114301
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