Colorimetric DNA-based assay for the specific detection and quantification of Ostreopsis cf. ovata and Ostreopsis cf. siamensis in the marine environment
Andree, Karl B.
O'Sullivan, Ciara K.
Ostreopsis is a toxic benthic dinoflagellate largely distributed worldwide in tropical and temperate areas. In the Mediterranean Sea, periodic summer blooms have been reported and have become a serious concern due to their direct impact on human health and the environment. Current microalgae identification is performed via light microscopy, which is time-consuming and is not able to differentiate among Ostreopsis species. Therefore, there is mature need for rapid, specific and easy-to-use detection tools. In this work, a colorimetric assay exploiting a combination of recombinase polymerase amplification (RPA) and a sandwich hybridisation assay was developed for O. cf. ovata and O. cf. siamensis detection and quantification. The specificity of the system was demonstrated by cross-reactivity experiments and calibration curves were successfully constructed using genomic DNA, achieving limits of detection of 10 and 14 pg/μL for O. cf. ovata and O. cf. siamensis, respectively. The assay was applied to the analysis of planktonic and benthic environmental samples from different sites of the Catalan coast. Species-specific DNA quantifications were in agreement with qPCR analysis, demonstrating the reliability of the colorimetric approach. Significant correlations were also obtained between DNA quantifications and light microscopy counts. The approach may be a valuable tool to provide timely warnings, facilitate monitoring activities or study population dynamics, and paves the way towards the development of in situ tools for the monitoring of harmful algal blooms.
63 - Agriculture and related sciences and techniques
Is part of
Toldrà, Anna, Carles Alcaraz, Karl B. Andree, Margarita Fernández-Tejedor, Jorge Diogène, Ioanis Katakis, Ciara K. O’Sullivan, and Mònica Campàs. 2019. "Colorimetric DNA-Based Assay For The Specific Detection And Quantification Of Ostreopsis Cf. Ovata And Ostreopsis Cf. Siamensis In The Marine Environment". Harmful Algae 84: 27-35. Elsevier BV. doi:10.1016/j.hal.2019.02.003.
Grant agreement number
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
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
Aigües Marines i Continentals
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