Development and Evaluation of a PCR Based Assay for Detection of the Toxic Dinoflagellate, <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis>(Graham) in Ballast Water and Environmental Samples |
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Authors: | Email author" target="_blank">Jawahar?G?PatilEmail author Rasanthi?M?Gunasekera Bruce?E?Deagle Nicholas?J?Bax Susan?I?Blackburn |
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Institution: | (1) CSIRO Marine Research, GPO Box 1538, Hobart, Tasmania, 7001, Australia |
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Abstract: | Gymnodinium catenatum is a bloom forming dinoflagellate that has been known to cause paralytic shellfish poisoning (PSP) in humans. It is being
reported with increased frequency around the world, with ballast water transport implicated as a primary vector that may have
contributed to its global spread. Major limitations to monitoring and management of its spread are the inability for early,
rapid, and accurate detection of G. catenatum in plankton samples. This study explored the feasibility of developing a PCR-based method for specific detection of G. catenatumin cultures and heterogeneous ballast water and environmental samples. Sequence comparison of the large sub unit (LSU) ribosomal
DNA locus of several strains and species of dinoflagellates allowed the design of G. catenatum specific PCR primers that are flanked by conserved regions. Assay specificity was validated through screening a range of
dinoflagellate cultures, including the morphologically similar and taxonomically closely related species G. nolleri. Amplification of the diagnostic PCR product from all the strains of G. catenatum but not from other species of dinoflagellates tested imply the species specificity of the assay. Sensitivity of the assay
to detect cysts in ballast water samples was established by simulated spiked experiments. The assay could detect G. catenatum in all ‘blank’ plankton samples that were spiked with five or more cysts. The assay was used to test environmental samples
collected from the Derwent river estuary, Tasmania. Based on the results we conclude that the assay may be utilized in large
scale screening of environmental and ballast water samples. |
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Keywords: | cyst dinoflagellate Gymnodinium catenatum introduced pest PCR detection |
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