Detection and quantification of the toxic marine microalgae Karlodinium veneficum and Karlodinium armiger using recombinase polymerase amplification and enzyme-linked oligonucleotide assay.

Detection and quantification of the toxic marine microalgae Karlodinium veneficum and Karlodinium armiger using recombinase polymerase amplification and enzyme-linked oligonucleotide assay.
Anal Chim Acta. 2018 Dec 18;1039:140-148
Authors: Toldrà A, Jauset-Rubio M, Andree KB, Fernández-Tejedor M, Diogène J, Katakis I, O'Sullivan CK, Campàs M
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.
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Source: Analytica Chimica Acta - October 18, 2018 Category: Chemistry Authors: Toldrà A, Jauset-Rubio M, Andree KB, Fernández-Tejedor M, Diogène J, Katakis I, O'Sullivan CK, Campàs M Tags: Anal Chim Acta Source Type: research