title: Molecular methods: chip assay and quantitative real-time PCR: in detecting hepatotoxic cyanobacteria.
abstract: Cyanobacterial mass occurrences are widespread and often contain hepatotoxic, i.e. microcystin- and nodularin-producing, species. Nowadays, detection of microcystin (mcy) and nodularin synthetase (nda) genes is widely used for the recognition of toxic cyanobacterial strains in environmental water samples. Chip assay presented here combines ligation detection reaction and hybridization on a universal microarray to detect and identify the mcyE/ndaF genes of five cyanobacterial genera specifically and sensitively. Thus, one chip assay can reveal the co-occurrence of several hepatotoxin producers. The presented quantitative real-time PCR method is used for the detection of either microcystin-producing Anabaena or Microcystis. Determination of the mcyE-gene copy numbers allows the identification of the dominant producer genus in the sample.
authors.affiliation: Division of Microbiology, Department of Food and Environmental Sciences, University of Helsinki, Helsinki, Finland.
authors.forename: Anne
authors.initials: A
authors.lastname: Rantala-Ylinen
authors.forename: Hanna
authors.initials: H
authors.lastname: Sipari
authors.forename: Kaarina
authors.initials: K
authors.lastname: Sivonen
mesh_terms: D017033:Anabaena / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000032:analysis* / Q000235:genetics; D015373:Bacterial Typing Techniques; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D018628:Gene Dosage; D005798:Genes, Bacterial; D046228:Microarray Analysis / Q000379:methods*; D052998:Microcystins / Q000032:analysis*; D046931:Microcystis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D010456:Peptides, Cyclic / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D052998:Microcystins; D010456:Peptides, Cyclic; D012336:RNA, Ribosomal, 16S; C063998:nodularin; C078588:microcystin