A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1
The Ostreid herpesvirus 1 species affects shellfish, contributing significantly to high economic losses during production. To counteract the threat related to mortality, there is a need for the development of novel point-of-care testing (POCT) that can be implemented in aquaculture production to pre...
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ftpubmed:oai:pubmedcentral.nih.gov:7555478 2023-05-15T15:58:35+02:00 A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 Zaczek-Moczydłowska, Maja A. Mohamed-Smith, Letitia Toldrà, Anna Hooper, Chantelle Campàs, Mònica Furones, M. Dolors Bean, Tim P. Campbell, Katrina 2020-09-09 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555478/ http://www.ncbi.nlm.nih.gov/pubmed/32917059 https://doi.org/10.3390/ijms21186605 en eng MDPI http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555478/ http://www.ncbi.nlm.nih.gov/pubmed/32917059 http://dx.doi.org/10.3390/ijms21186605 © 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). CC-BY Int J Mol Sci Article Text 2020 ftpubmed https://doi.org/10.3390/ijms21186605 2020-10-25T00:33:05Z The Ostreid herpesvirus 1 species affects shellfish, contributing significantly to high economic losses during production. To counteract the threat related to mortality, there is a need for the development of novel point-of-care testing (POCT) that can be implemented in aquaculture production to prevent disease outbreaks. In this study, a simple, rapid and specific colorimetric loop-mediated isothermal amplification (LAMP) assay has been developed for the detection of Ostreid herpesvirus 1 (OsHV-1) and its variants infecting Crassostrea gigas (C. gigas). The LAMP assay has been optimized to use hydroxynaphthol blue (HNB) for visual colorimetric distinction of positive and negative templates. The effect of an additional Tte UvrD helicase enzyme used in the reaction was also evaluated with an improved reaction time of 10 min. Additionally, this study provides a robust workflow for optimization of primers for uncultured viruses using designed target plasmid when DNA availability is limited. Text Crassostrea gigas PubMed Central (PMC) International Journal of Molecular Sciences 21 18 6605 |
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Article Zaczek-Moczydłowska, Maja A. Mohamed-Smith, Letitia Toldrà, Anna Hooper, Chantelle Campàs, Mònica Furones, M. Dolors Bean, Tim P. Campbell, Katrina A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
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Article |
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The Ostreid herpesvirus 1 species affects shellfish, contributing significantly to high economic losses during production. To counteract the threat related to mortality, there is a need for the development of novel point-of-care testing (POCT) that can be implemented in aquaculture production to prevent disease outbreaks. In this study, a simple, rapid and specific colorimetric loop-mediated isothermal amplification (LAMP) assay has been developed for the detection of Ostreid herpesvirus 1 (OsHV-1) and its variants infecting Crassostrea gigas (C. gigas). The LAMP assay has been optimized to use hydroxynaphthol blue (HNB) for visual colorimetric distinction of positive and negative templates. The effect of an additional Tte UvrD helicase enzyme used in the reaction was also evaluated with an improved reaction time of 10 min. Additionally, this study provides a robust workflow for optimization of primers for uncultured viruses using designed target plasmid when DNA availability is limited. |
format |
Text |
author |
Zaczek-Moczydłowska, Maja A. Mohamed-Smith, Letitia Toldrà, Anna Hooper, Chantelle Campàs, Mònica Furones, M. Dolors Bean, Tim P. Campbell, Katrina |
author_facet |
Zaczek-Moczydłowska, Maja A. Mohamed-Smith, Letitia Toldrà, Anna Hooper, Chantelle Campàs, Mònica Furones, M. Dolors Bean, Tim P. Campbell, Katrina |
author_sort |
Zaczek-Moczydłowska, Maja A. |
title |
A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
title_short |
A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
title_full |
A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
title_fullStr |
A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
title_full_unstemmed |
A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification for the Robust Detection of the Ostreid herpesvirus 1 |
title_sort |
single-tube hnb-based loop-mediated isothermal amplification for the robust detection of the ostreid herpesvirus 1 |
publisher |
MDPI |
publishDate |
2020 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555478/ http://www.ncbi.nlm.nih.gov/pubmed/32917059 https://doi.org/10.3390/ijms21186605 |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
Int J Mol Sci |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7555478/ http://www.ncbi.nlm.nih.gov/pubmed/32917059 http://dx.doi.org/10.3390/ijms21186605 |
op_rights |
© 2020 by the authors. 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 (http://creativecommons.org/licenses/by/4.0/). |
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CC-BY |
op_doi |
https://doi.org/10.3390/ijms21186605 |
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International Journal of Molecular Sciences |
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21 |
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18 |
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6605 |
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1766394343740932096 |