Detection and quantification of the toxic marine microalgae Karlodinium veneficum and Karlodinium armiger using recombinase polymerase amplification and enzyme-linked oligonucleotide assay
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Author
Toldrà, Anna
Jauset-Rubio, Míriam
Andree, Karl B.
Katakis, Ioanis
O'Sullivan, Ciara K.
Publication date
2018-07-28ISSN
0003-2670
Abstract
Karlodinium is a dinoflagellate responsible for fish-killing events worldwide. In Alfacs Bay (NW Mediterranean Sea), the presence of two Karlodinium species (K. veneficum and K. armiger) with different toxicities has been reported. This work presents a method that combines recombinase polymerase amplification (RPA) with an enzyme-linked oligonucleotide assay (ELONA) to identify, discriminate and quantify these two species. The system was characterised using synthetic DNA and genomic DNA, and the specificity was confirmed by cross-reactivity experiments. Calibration curves were constructed using 10-fold dilutions of cultured cells, attaining a limit of detection of around 50,000 cells/L, far below the Karlodinium spp. alert threshold (200,000 cells/L). Finally, the assay was applied to spiked seawater samples, showing an excellent correlation with the spiking levels and light microscopy counts. This approach is more rapid, specific and user-friendly than traditional microscopy techniques, and shows great promise for the surveillance and management of harmful algal blooms.
Document Type
Article
Document version
Accepted version
Language
English
Subject (CDU)
63 - Agriculture and related sciences and techniques
Pages
19
Publisher
Elsevier
Is part of
Analytica Chimica Acta
Citation
Toldrà, Anna, Míriam Jauset-Rubio, Karl B. Andree, Margarita Fernández-Tejedor, Jorge Diogène, Ioanis Katakis, Ciara K. O'Sullivan, and Mònica Campàs. 2018. "Detection And Quantification Of The Toxic Marine Microalgae Karlodinium Veneficum And Karlodinium Armiger Using Recombinase Polymerase Amplification And Enzyme-Linked Oligonucleotide Assay". Analytica Chimica Acta 1039: 140-148. Elsevier BV. doi:10.1016/j.aca.2018.07.057.
Grant agreement number
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2017-87946-C2-2-R/ES/DISPOSITIVOS BIOANALITICOS PARA LA DETECCION RAPIDA Y RENTABLE DE GAMBIERDISCUS Y CIGUATOXINAS/CIGUASENSING
MINECO/Programa Estatal de I+D+I orientada a los retos de la sociedad/BIO2014-56024-C2-2-R/ES/MICROSISTEMAS PARA LA DETECCION RAPIDA, FIABLE Y RENTABLE DE MICROALGAS TOXICAS IN SITU Y A TIEMPO REAL/SEASENSING
Program
Aigües Marines i Continentals
Aqüicultura
This item appears in the following Collection(s)
- ARTICLES CIENTÍFICS [2820]
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by-nc-nd/4.0/