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Species-specific real-time PCR cell number quantification of the bloom forming cyanobacterium Planktothrix agardhii

2012-09Journal article Restricted access
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dc.contributor.authorChurro, Catarina
dc.contributor.authorPereira, Paulo
dc.contributor.authorVasconcelos, Vitor
dc.contributor.authorValério, Elisabete
dc.date.accessioned2012-11-26T12:50:37Z
dc.date.available2012-11-26T12:50:37Z
dc.date.issued2012-09
dc.description.abstractA species-specific method to detect and quantify Planktothrix agardhii was developed by combining the SYBR Green I real-time polymerase chain reaction technique with a simplified DNA extraction procedure for standard curve preparation. Newly designed PCR primers were used to amplify a specific fragment within the rpoC1 gene. Since this gene exists in single copy in the genome, it allows the direct achievement of cell concentrations. The cell concentration determined by real-time PCR showed a linear correlation with the cell concentration determined from direct microscopic counts. The detection limit for cell quantification of the method was 8 cells μL−1, corresponding to 32 cells per reaction. Furthermore, the real-time qPCR method described in this study allowed a successful quantification of P. agardhii from environmental water samples, showing that this protocol is an accurate and economic tool for a rapid absolute quantification of the potentially toxic cyanobacterium P. agardhii.por
dc.identifier.citationArch Microbiol. 2012 Sep;194(9):749-57. Epub 2012 Apr 7.por
dc.identifier.issn0302-8933
dc.identifier.otherdoi: 10.1007/s00203-012-0809-y
dc.identifier.urihttp://hdl.handle.net/10400.18/1119
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherSpringer Verlagpor
dc.relationPPCDT/ AMB/67075/2006 and PTDC/AAC-CLI/116122/2009por
dc.relation.publisherversionhttp://link.springer.com/article/10.1007%2Fs00203-012-0809-ypor
dc.subjectCyanobacteriapor
dc.subjectPlanktothrix Agardhiipor
dc.subjectReal-time qPCRpor
dc.subjectrpoC1 Genepor
dc.subjectÁgua e Solopor
dc.titleSpecies-specific real-time PCR cell number quantification of the bloom forming cyanobacterium Planktothrix agardhiipor
dc.typejournal article
dspace.entity.typePublication
oaire.citation.endPage757por
oaire.citation.startPage749por
oaire.citation.titleArchives of Microbiologypor
oaire.citation.volume194por
rcaap.rightsrestrictedAccesspor
rcaap.typearticlepor

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