A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries

We developed a NitroPure Nitrocellulose (NPN) membrane-based method for sampling and storing grapevine sap for grapevine virus detection. We devised an efficient nucleic acid extraction method for the NPN membrane, resulting in 100% amplification success for grapevine leafroll-associated virus 2 (GL...

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Autores principales: Nita, Mizuho, Jones, Taylor, McHenry, Diana, Bush, Elizabeth, Oliver, Charlotte, Kawaguchi, Akira, Nita, Akiko, Katori, Miyuki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10612103/
https://www.ncbi.nlm.nih.gov/pubmed/37896878
http://dx.doi.org/10.3390/v15102102
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author Nita, Mizuho
Jones, Taylor
McHenry, Diana
Bush, Elizabeth
Oliver, Charlotte
Kawaguchi, Akira
Nita, Akiko
Katori, Miyuki
author_facet Nita, Mizuho
Jones, Taylor
McHenry, Diana
Bush, Elizabeth
Oliver, Charlotte
Kawaguchi, Akira
Nita, Akiko
Katori, Miyuki
author_sort Nita, Mizuho
collection PubMed
description We developed a NitroPure Nitrocellulose (NPN) membrane-based method for sampling and storing grapevine sap for grapevine virus detection. We devised an efficient nucleic acid extraction method for the NPN membrane, resulting in 100% amplification success for grapevine leafroll-associated virus 2 (GLRaV2) and 3 (GLRaV3), grapevine rupestris stem pitting-associated virus (GRSPaV), grapevine virus A, grapevine virus B, and grapevine red blotch virus (GRBV). This method also allowed the storage of recoverable nucleic acid for 18 months at room temperature. We created a sampling kit to survey GLRaV2, GLRaV3, and GRBV in Japanese vineyards. We tested the kits in the field in 2018 and then conducted mail-in surveys in 2020–2021. The results showed a substantial prevalence of GLRaV3, with 48.5% of 132 sampled vines being positive. On the other hand, only 3% of samples tested positive for GLRaV2 and none for GRBV.
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spelling pubmed-106121032023-10-29 A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries Nita, Mizuho Jones, Taylor McHenry, Diana Bush, Elizabeth Oliver, Charlotte Kawaguchi, Akira Nita, Akiko Katori, Miyuki Viruses Article We developed a NitroPure Nitrocellulose (NPN) membrane-based method for sampling and storing grapevine sap for grapevine virus detection. We devised an efficient nucleic acid extraction method for the NPN membrane, resulting in 100% amplification success for grapevine leafroll-associated virus 2 (GLRaV2) and 3 (GLRaV3), grapevine rupestris stem pitting-associated virus (GRSPaV), grapevine virus A, grapevine virus B, and grapevine red blotch virus (GRBV). This method also allowed the storage of recoverable nucleic acid for 18 months at room temperature. We created a sampling kit to survey GLRaV2, GLRaV3, and GRBV in Japanese vineyards. We tested the kits in the field in 2018 and then conducted mail-in surveys in 2020–2021. The results showed a substantial prevalence of GLRaV3, with 48.5% of 132 sampled vines being positive. On the other hand, only 3% of samples tested positive for GLRaV2 and none for GRBV. MDPI 2023-10-17 /pmc/articles/PMC10612103/ /pubmed/37896878 http://dx.doi.org/10.3390/v15102102 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Nita, Mizuho
Jones, Taylor
McHenry, Diana
Bush, Elizabeth
Oliver, Charlotte
Kawaguchi, Akira
Nita, Akiko
Katori, Miyuki
A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title_full A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title_fullStr A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title_full_unstemmed A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title_short A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries
title_sort nitropure nitrocellulose membrane-based grapevine virus sampling kit: development and deployment to survey japanese vineyards and nurseries
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10612103/
https://www.ncbi.nlm.nih.gov/pubmed/37896878
http://dx.doi.org/10.3390/v15102102
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