Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae
The European oyster Ostrea edulis played a key role in the North Sea by providing several ecosystem functions and services. Today, O. edulis is classified as severely degraded or functionally extinct in Europe. Marine conservation is focusing on biogenic reef restoration, namely the restoration of O...
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EDP Sciences
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Online Access: | https://doi.org/10.1051/alr/2021023 https://doaj.org/article/28f5d1e60df84c218f03906c4a578bd6 |
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ftdoajarticles:oai:doaj.org/article:28f5d1e60df84c218f03906c4a578bd6 2023-10-09T21:50:56+02:00 Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae Benkens Andreas Buchholz Claudia Pogoda Bernadette Harms Carsten Georg 2021-01-01T00:00:00Z https://doi.org/10.1051/alr/2021023 https://doaj.org/article/28f5d1e60df84c218f03906c4a578bd6 EN eng EDP Sciences https://www.alr-journal.org/articles/alr/full_html/2021/01/alr210051/alr210051.html https://doaj.org/toc/1765-2952 1765-2952 doi:10.1051/alr/2021023 https://doaj.org/article/28f5d1e60df84c218f03906c4a578bd6 Aquatic Living Resources, Vol 34, p 23 (2021) nucleic acid lateral flow immunoassay oyster restoration larvae detection european oyster point-of-care detection biogenic reef restoration Aquaculture. Fisheries. Angling SH1-691 article 2021 ftdoajarticles https://doi.org/10.1051/alr/2021023 2023-09-10T00:44:35Z The European oyster Ostrea edulis played a key role in the North Sea by providing several ecosystem functions and services. Today, O. edulis is classified as severely degraded or functionally extinct in Europe. Marine conservation is focusing on biogenic reef restoration, namely the restoration of O. edulis in Natura 2000 sites of the North Sea. The identification of oyster larvae related to natural spatfalls of restored reefs and monitoring of larval drift is a key aspect of marine protected area management. Morphological identification and distinction from other abundant bivalve larvae using microscopy is difficult. Existing molecular biological methods are expensive and bound to stationary laboratory equipment, or are inadequate in the visualization. In this study, we identified nucleic acid lateral flow immunoassay (NALFIA), a well-established tool in human pathogen diagnostics, as an efficient approach for point-of-care (POC) testing in marine monitoring. Based on the genetic sequence of the mitochondrial cytochrome b of O. edulis, forward and reverse primers were developed. The reverse primer was labelled with fluorescent dye FITC, forward primer with biotin. Reaction on the lateral flow stripe could be realized with a single O. edulis larva in direct PCR with multiplex primers in a portable PCR-cycler. The established NALFIA system can distinguish O. edulis larvae from Crassostrea gigas and Mytilus edulis larvae, respectively. This method offers new approaches in POC testing in marine research and monitoring. It gives quick and clear results, is inexpensive, and could be easily adapted to other species of interest. Article in Journal/Newspaper Crassostrea gigas Directory of Open Access Journals: DOAJ Articles Stripe ENVELOPE(9.914,9.914,63.019,63.019) Aquatic Living Resources 34 23 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
nucleic acid lateral flow immunoassay oyster restoration larvae detection european oyster point-of-care detection biogenic reef restoration Aquaculture. Fisheries. Angling SH1-691 |
spellingShingle |
nucleic acid lateral flow immunoassay oyster restoration larvae detection european oyster point-of-care detection biogenic reef restoration Aquaculture. Fisheries. Angling SH1-691 Benkens Andreas Buchholz Claudia Pogoda Bernadette Harms Carsten Georg Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
topic_facet |
nucleic acid lateral flow immunoassay oyster restoration larvae detection european oyster point-of-care detection biogenic reef restoration Aquaculture. Fisheries. Angling SH1-691 |
description |
The European oyster Ostrea edulis played a key role in the North Sea by providing several ecosystem functions and services. Today, O. edulis is classified as severely degraded or functionally extinct in Europe. Marine conservation is focusing on biogenic reef restoration, namely the restoration of O. edulis in Natura 2000 sites of the North Sea. The identification of oyster larvae related to natural spatfalls of restored reefs and monitoring of larval drift is a key aspect of marine protected area management. Morphological identification and distinction from other abundant bivalve larvae using microscopy is difficult. Existing molecular biological methods are expensive and bound to stationary laboratory equipment, or are inadequate in the visualization. In this study, we identified nucleic acid lateral flow immunoassay (NALFIA), a well-established tool in human pathogen diagnostics, as an efficient approach for point-of-care (POC) testing in marine monitoring. Based on the genetic sequence of the mitochondrial cytochrome b of O. edulis, forward and reverse primers were developed. The reverse primer was labelled with fluorescent dye FITC, forward primer with biotin. Reaction on the lateral flow stripe could be realized with a single O. edulis larva in direct PCR with multiplex primers in a portable PCR-cycler. The established NALFIA system can distinguish O. edulis larvae from Crassostrea gigas and Mytilus edulis larvae, respectively. This method offers new approaches in POC testing in marine research and monitoring. It gives quick and clear results, is inexpensive, and could be easily adapted to other species of interest. |
format |
Article in Journal/Newspaper |
author |
Benkens Andreas Buchholz Claudia Pogoda Bernadette Harms Carsten Georg |
author_facet |
Benkens Andreas Buchholz Claudia Pogoda Bernadette Harms Carsten Georg |
author_sort |
Benkens Andreas |
title |
Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
title_short |
Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
title_full |
Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
title_fullStr |
Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
title_full_unstemmed |
Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae |
title_sort |
direct multiplex pcr-nalfia to inform marine conservation: use of an innovative diagnostic tool for the detection of ostrea edulis larvae |
publisher |
EDP Sciences |
publishDate |
2021 |
url |
https://doi.org/10.1051/alr/2021023 https://doaj.org/article/28f5d1e60df84c218f03906c4a578bd6 |
long_lat |
ENVELOPE(9.914,9.914,63.019,63.019) |
geographic |
Stripe |
geographic_facet |
Stripe |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
Aquatic Living Resources, Vol 34, p 23 (2021) |
op_relation |
https://www.alr-journal.org/articles/alr/full_html/2021/01/alr210051/alr210051.html https://doaj.org/toc/1765-2952 1765-2952 doi:10.1051/alr/2021023 https://doaj.org/article/28f5d1e60df84c218f03906c4a578bd6 |
op_doi |
https://doi.org/10.1051/alr/2021023 |
container_title |
Aquatic Living Resources |
container_volume |
34 |
container_start_page |
23 |
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1779314022896631808 |