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
; 1039: 140-148, 2018 Dec 18.
Article
en En
| MEDLINE
| ID: mdl-30322545
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 50,000â¯cells/L, far below the Karlodinium spp. alert threshold (200,000â¯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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Oligonucleótidos
/
Ensayo de Inmunoadsorción Enzimática
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Microalgas
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Reacción en Cadena en Tiempo Real de la Polimerasa
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Toxinas Marinas
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Anal Chim Acta
Año:
2018
Tipo del documento:
Article
País de afiliación:
España