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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ftawi:oai:epic.awi.de:55648 2024-09-15T18:03:16+00:00 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 2021-10-15 https://epic.awi.de/id/eprint/55648/ https://www.alr-journal.org/articles/alr/abs/2021/01/alr210051/alr210051.html https://hdl.handle.net/10013/epic.2c1d36ea-f4c1-4382-90ee-881623ca399a unknown EDP SCIENCES S A Benkens, A. orcid:0000-0001-7943-7843 , Buchholz, C. orcid:0000-0002-5055-1586 , Pogoda, B. orcid:0000-0003-3997-426X and Harms, C. G. (2021) Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae , Aquatic Living Resources, 34 (23), pp. 1-16 . doi:10.1051/alr/2021023 <https://doi.org/10.1051/alr%2F2021023> , hdl:10013/epic.2c1d36ea-f4c1-4382-90ee-881623ca399a EPIC3Aquatic Living Resources, EDP SCIENCES S A, 34(23), pp. 1-16, ISSN: 0990-7440 Article peerRev 2021 ftawi https://doi.org/10.1051/alr/2021023 2024-06-24T04:27:29Z 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 Alfred Wegener Institute for Polar- and Marine Research (AWI): ePIC (electronic Publication Information Center) Aquatic Living Resources 34 23 |
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Alfred Wegener Institute for Polar- and Marine Research (AWI): ePIC (electronic Publication Information Center) |
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ftawi |
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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 |
spellingShingle |
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 |
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 S A |
publishDate |
2021 |
url |
https://epic.awi.de/id/eprint/55648/ https://www.alr-journal.org/articles/alr/abs/2021/01/alr210051/alr210051.html https://hdl.handle.net/10013/epic.2c1d36ea-f4c1-4382-90ee-881623ca399a |
genre |
Crassostrea gigas |
genre_facet |
Crassostrea gigas |
op_source |
EPIC3Aquatic Living Resources, EDP SCIENCES S A, 34(23), pp. 1-16, ISSN: 0990-7440 |
op_relation |
Benkens, A. orcid:0000-0001-7943-7843 , Buchholz, C. orcid:0000-0002-5055-1586 , Pogoda, B. orcid:0000-0003-3997-426X and Harms, C. G. (2021) Direct multiplex PCR-NALFIA to inform marine conservation: Use of an innovative diagnostic tool for the detection of Ostrea edulis larvae , Aquatic Living Resources, 34 (23), pp. 1-16 . doi:10.1051/alr/2021023 <https://doi.org/10.1051/alr%2F2021023> , hdl:10013/epic.2c1d36ea-f4c1-4382-90ee-881623ca399a |
op_doi |
https://doi.org/10.1051/alr/2021023 |
container_title |
Aquatic Living Resources |
container_volume |
34 |
container_start_page |
23 |
_version_ |
1810440788467777536 |