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Multiplex PCR for detection of microcystins-producing cyanobacteria from freshwater samples

2010-06Journal article Restricted access
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dc.contributor.authorValério, Elisabete
dc.contributor.authorChambel, Lélia
dc.contributor.authorPaulino, Sérgio
dc.contributor.authorFaria, Natália
dc.contributor.authorPereira, Paulo
dc.contributor.authorTenreiro, Rogério
dc.date.accessioned2011-09-19T11:44:13Z
dc.date.available2011-09-19T11:44:13Z
dc.date.issued2010-06
dc.description.abstractThe aim of this study was to develop a PCR-based method of gene-directed multiplex PCR to rapidly identify microcystins producing cyanobacteria, regardless of their taxa, that could be applied in routine freshwater monitoring. Instead of using the amplification of only one or two mcy gene fragments, a multiplex PCR that simultaneously amplifies mcyA-cd, mcyAB, and mcyB fragments of the microcystin gene cluster was validated with DNA from 124 cyanobacterial isolates and applied in 37 environmental samples. The toxicological status of the isolates was assessed by high-performance liquid chromatography also used as the "gold standard" for the evaluation of multiplex mcy genes-based PCR, where a sensitivity of 92.3% and a specificity of 100% have been obtained. For the environmental samples, a rapid protocol for their direct use in the PCR reaction has been developed and, by using ELISA results as "gold standard" for the presence of microcystins in these samples, a sensitivity of 80% and a specificity of 100% were achieved, showing that this multiplex PCR test is a rapid, reliable, and economical way of assessing the microcystin-producing potential of cyanobacteria in freshwaters, regardless of their taxa or microcystins variant produced.por
dc.identifier.citationEnviron Toxicol. 2010 Jun;25(3):251-60. Epub 2009 Jun 1por
dc.identifier.issn1520-4081
dc.identifier.otherdoi:10.1002/tox.20502
dc.identifier.urihttp://hdl.handle.net/10400.18/191
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherWiley-Blackwellpor
dc.relation.publisherversionhttp://onlinelibrary.wiley.com/doi/10.1002/tox.20502/pdfpor
dc.subjectCyanobacteriapor
dc.subjectMicrocystinspor
dc.subjectMultiplex PCRpor
dc.subjectEnvironmental samplespor
dc.subjectBloomspor
dc.subjectÁgua e Solopor
dc.titleMultiplex PCR for detection of microcystins-producing cyanobacteria from freshwater samplespor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage260por
oaire.citation.startPage251por
oaire.citation.titleEnvironmental Toxicologypor
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor

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