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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ftirta:oai:repositori.irta.cat:20.500.12327/938 2023-07-02T03:32:03+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 Producció Animal Aigües Marines i Continentals Aqüicultura 2020-09-09 15 application/pdf https://hdl.handle.net/20.500.12327/938 https://doi.org/10.3390/ijms21186605 eng eng MDPI International Journal of Molecular Sciences EC/H2020/678589/EU/Preventing and mitigating farmed bivalve diseases/VIVALDI Zaczek-Moczydłowska, Maja A., Letitia Mohamed-Smith, Anna Toldrà, Chantelle Hooper, Mònica Campàs, M. Dolors Furones, Tim P. Bean, and Katrina Campbell. 2020. "A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification For The Robust Detection Of The Ostreid Herpesvirus 1". International Journal Of Molecular Sciences 21 (18): 6605. doi:10.3390/ijms21186605. 1661-6596 http://hdl.handle.net/20.500.12327/938 https://doi.org/10.3390/ijms21186605 Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess 639 info:eu-repo/semantics/article 2020 ftirta https://doi.org/20.500.12327/93810.3390/ijms21186605 2023-06-13T23:20:02Z 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 herpesvirus1 (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. info:eu-repo/semantics/publishedVersion Article in Journal/Newspaper Crassostrea gigas IRTA Pubpro (Institute of Agrifood Research and Technology) International Journal of Molecular Sciences 21 18 6605 |
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English |
topic |
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639 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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639 |
description |
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 herpesvirus1 (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. info:eu-repo/semantics/publishedVersion |
author2 |
Producció Animal Aigües Marines i Continentals Aqüicultura |
format |
Article in Journal/Newspaper |
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 |
https://hdl.handle.net/20.500.12327/938 https://doi.org/10.3390/ijms21186605 |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_relation |
International Journal of Molecular Sciences EC/H2020/678589/EU/Preventing and mitigating farmed bivalve diseases/VIVALDI Zaczek-Moczydłowska, Maja A., Letitia Mohamed-Smith, Anna Toldrà, Chantelle Hooper, Mònica Campàs, M. Dolors Furones, Tim P. Bean, and Katrina Campbell. 2020. "A Single-Tube HNB-Based Loop-Mediated Isothermal Amplification For The Robust Detection Of The Ostreid Herpesvirus 1". International Journal Of Molecular Sciences 21 (18): 6605. doi:10.3390/ijms21186605. 1661-6596 http://hdl.handle.net/20.500.12327/938 https://doi.org/10.3390/ijms21186605 |
op_rights |
Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/20.500.12327/93810.3390/ijms21186605 |
container_title |
International Journal of Molecular Sciences |
container_volume |
21 |
container_issue |
18 |
container_start_page |
6605 |
_version_ |
1770271527130890240 |