Please use this identifier to cite or link to this item:
DC FieldValue
dc.titleDevelopment of semi-quantitative pcr assays for the detection and enumeration of gambierdiscus species (gonyaulacales, dinophyceae)
dc.contributor.authorVandersea, M.W.
dc.contributor.authorKibler, S.R.
dc.contributor.authorHolland, W.C.
dc.contributor.authorTester, P.A.
dc.contributor.authorSchultz, T.F.
dc.contributor.authorFaust, M.A.
dc.contributor.authorHolmes, M.J.
dc.contributor.authorChinain, M.
dc.contributor.authorWayne Litaker, R.
dc.identifier.citationVandersea, M.W., Kibler, S.R., Holland, W.C., Tester, P.A., Schultz, T.F., Faust, M.A., Holmes, M.J., Chinain, M., Wayne Litaker, R. (2012-08). Development of semi-quantitative pcr assays for the detection and enumeration of gambierdiscus species (gonyaulacales, dinophyceae). Journal of Phycology 48 (4) : 902-915. ScholarBank@NUS Repository.
dc.description.abstractCiguatera fish poisoning (CFP) is a serious health problem in tropical regions and is caused by the bioaccumulation of lipophilic toxins produced by dinoflagellates in the genus Gambierdiscus. Gambierdiscus species are morphologically similar and are difficult to distinguish from one another even when using scanning electron microscopy. Improved identification and detection methods that are sensitive and rapid are needed to identify toxic species and investigate potential distribution and abundance patterns in relation to incidences of CFP. This study presents the first species-specific, semi-quantitative polymerase chain reaction (qPCR) assays that can be used to address these questions. These assays are specific for five Gambierdiscus species and one undescribed ribotype. The assays utilized a SYBR green format and targeted unique sequences found within the SSU, ITS, and the D1/D3 LSU ribosomal domains. Standard curves were constructed using known concentrations of cultured cells and 10-fold serial dilutions of rDNA PCR amplicons containing the target sequence for each specific assay. Assay sensitivity and accuracy were tested using DNA extracts purified from known concentrations of multiple Gambierdiscus species. The qPCR assays were used to assess Gambierdiscus species diversity and abundance in samples collected from nearshore areas adjacent to Ft. Pierce and Jupiter, Florida USA. The results indicated that the practical limit of detection for each assay was 10 cells per sample. Most interestingly, the qPCR analysis revealed that as many as four species of Gambierdiscus were present in a single macrophyte sample. © 2012 Phycological Society of America.
dc.subjectGambierdiscus belizeanus
dc.subjectGambierdiscus caribaeus
dc.subjectGambierdiscus carolinianus
dc.subjectGambierdiscus carpenteri
dc.subjectGambierdiscus ribotype 2
dc.subjectGambierdiscus ruetzleri
dc.subjectSYBR green
dc.description.sourcetitleJournal of Phycology
Appears in Collections:Staff Publications

Show simple item record
Files in This Item:
There are no files associated with this item.

Google ScholarTM



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.