Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples

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dc.contributor.advisorO'Ryan, Cen_ZA
dc.contributor.advisorSebastián, C Ruizen_ZA
dc.contributor.authorVan Helmond, Zen_ZA
dc.date.accessioned2014-08-13T14:10:28Z
dc.date.available2014-08-13T14:10:28Z
dc.date.issued2004en_ZA
dc.descriptionIncludes bibliographical references (leaves. 23-28).en_ZA
dc.description.abstractThe frequency and geographical range of hannful algal blooms (HABs) are believed to be on the increase, with adverse affects on marine and human health making the implementation of stringent controls governmg monitoring programmes commonplace. The South African monitoring programme was established in 1989 and relies upon microscopic identification of HAB species. Microscopic identification is labour-intensive, requiring a high level of taxonomic expertise, and could be considered impractical for routine monitoring where analysis of large numbers of samples is required. Novel monitoring techniques, focusing mainly on probe technology, are being developed for rapid, unequivocal identification and enumeration of HAB species. In this study, a triplex peR assay, incorporating a genus-specific ribosomal DNA primer designed from phylogenetic studies on local Alexandrium populations, was optimised for application to environmental samples and tested against natural assemblages containing Alexandrium minutum. Specific positive results were consistently generated for samples containing A. minutum. Samples absent of A. minuturn cells did not generate the Alexandrium-specific amplicon. The absolute detection limit of 440 A. minutum cells r1 for this assay was established. Effects of non-target cells on the sensitivity of the assay were also investigated: although a decrease in sensitivity was found, A. minutum cells could still be detected in the presence of 100 times more non-target cells. This assay has been shown to be a useful tool for unequivocal identification of A. minutum cells within local environmental samples.en_ZA
dc.identifier.apacitationVan Helmond, Z. (2004). <i>Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Biological Sciences. Retrieved from http://hdl.handle.net/11427/6197en_ZA
dc.identifier.chicagocitationVan Helmond, Z. <i>"Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Biological Sciences, 2004. http://hdl.handle.net/11427/6197en_ZA
dc.identifier.citationVan Helmond, Z. 2004. Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Van Helmond, Z AB - The frequency and geographical range of hannful algal blooms (HABs) are believed to be on the increase, with adverse affects on marine and human health making the implementation of stringent controls governmg monitoring programmes commonplace. The South African monitoring programme was established in 1989 and relies upon microscopic identification of HAB species. Microscopic identification is labour-intensive, requiring a high level of taxonomic expertise, and could be considered impractical for routine monitoring where analysis of large numbers of samples is required. Novel monitoring techniques, focusing mainly on probe technology, are being developed for rapid, unequivocal identification and enumeration of HAB species. In this study, a triplex peR assay, incorporating a genus-specific ribosomal DNA primer designed from phylogenetic studies on local Alexandrium populations, was optimised for application to environmental samples and tested against natural assemblages containing Alexandrium minutum. Specific positive results were consistently generated for samples containing A. minutum. Samples absent of A. minuturn cells did not generate the Alexandrium-specific amplicon. The absolute detection limit of 440 A. minutum cells r1 for this assay was established. Effects of non-target cells on the sensitivity of the assay were also investigated: although a decrease in sensitivity was found, A. minutum cells could still be detected in the presence of 100 times more non-target cells. This assay has been shown to be a useful tool for unequivocal identification of A. minutum cells within local environmental samples. DA - 2004 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2004 T1 - Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples TI - Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples UR - http://hdl.handle.net/11427/6197 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/6197
dc.identifier.vancouvercitationVan Helmond Z. Initial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samples. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Biological Sciences, 2004 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/6197en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDepartment of Biological Sciencesen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherZoologyen_ZA
dc.titleInitial assessment of triplex PCR assay application for detection of toxic dinoflagellates, Alexandrium species (Dinophyceae), in environmental samplesen_ZA
dc.typeMaster Thesis
dc.type.qualificationlevelMasters
dc.type.qualificationnameMScen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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