title: A new quantitative PCR assay for the detection of hepatotoxigenic cyanobacteria.
abstract: Toxin-producing cyanobacteria are a worldwide threat to both human and animal health. The hepatotoxins microcystin and nodularin are the most commonly occurring toxins produced by bloom-forming cyanobacteria. They are cyclic peptides that are synthesized nonribosomally by a multienzyme complexes encoded within the microcystin (mcyS) and nodularin (ndaS) synthetase gene clusters. Early detection of potentially toxic blooms would allow for pre-emptive action to reduce consumer exposure to cyanotoxins. We have developed a quantitative PCR (qPCR) assay based on SYBR-green chemistry for the detection of potentially hepatotoxic cyanobacteria spanning all known microcystin and nodularin producing taxa using primers specifically targeting mcyE and ndaF. The qPCR assay was validated against previously analyzed cyanobacterial bloom samples. Whole cell qPCR using cultured M. aeruginosa PCC7806 and non-toxic M. aeruginosa UTEX2386 had a sensitivity of 1000 cells ml⁻¹. In summary, we have developed a robust and sensitive molecular method for the detection and quantification of hepatotoxigenic cyanobacteria in bloom samples. This technology offers several advantages over traditional and contemporary testing protocols currently used to assess water quality.
authors.affiliation: School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney 2052, Australia.
authors.forename: J
authors.initials: J
authors.lastname: Al-Tebrineh
authors.forename: M M
authors.initials: MM
authors.lastname: Gehringer
authors.forename: R
authors.initials: R
authors.lastname: Akcaalan
authors.forename: B A
authors.initials: BA
authors.lastname: Neilan
mesh_terms: D001427:Bacterial Toxins / Q000032:analysis* / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004784:Environmental Monitoring; D057097:Harmful Algal Bloom; D052998:Microcystins / Q000032:analysis* / Q000235:genetics; D046932:Nodularia / Q000235:genetics / Q000302:isolation & purification*; D010456:Peptides, Cyclic / Q000032:analysis* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D014871:Water Microbiology
chemical_list: D001427:Bacterial Toxins; D004269:DNA, Bacterial; D052998:Microcystins; D010456:Peptides, Cyclic; C063998:nodularin