Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria.
Water bodies are routinely monitored for the presence of potentially toxic cyanobacteria; however, the methodology for confirming toxicity is currently complex and expensive. Here we describe the application of gene-based technology to rapidly identify cylindrospermopsin-producing cyanobacteria, specifically, Cylindrospermopsis raciborskii. A multiplex polymerase chain reaction (PCR) test was developed that simultaneously identified polyketide synthase (pks) and peptide synthetase (ps) determinants associated with cylindrospermopsin production and distinguished C. raciborskii from other cylindrospermopsin-producing cyanobacteria of the species Anabaena bergii and Aphanizomenon ovalisporum, by targeting the rpoC1 gene. Twenty-one C. raciborskii, 5 A. bergii, 10 Aph. ovalisporum isolates and 3 environmental samples all yielded PCR results consistent with their toxicological status, as assessed by high-performance liquid chromatography coupled to mass spectrometry or matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, and C. raciborskii was always correctly identified. The PCR test is a rapid, reliable, and economical way of assessing the toxic potential of cyanobacterial blooms formed by these organisms.
Fergusson
Kim M
KM
Cooperative Research Centre for Water Quality and Treatment, Australian Water Quality Centre, SA Water Corporation, Private Mail Bag 3, Salisbury, South Australia 5108, Australia.
Saint
Christopher P
CP
D000470:Alkaloids; D001427:Bacterial Toxins; D002851:Chromatography, High Pressure Liquid; D000458:Cyanobacteria / Q000235:genetics* / Q000472:pathogenicity*; D000087522:Cyanobacteria Toxins; D004269:DNA, Bacterial / Q000032:analysis*; D004784:Environmental Monitoring; D009097:Multienzyme Complexes / Q000032:analysis / Q000235:genetics*; D010453:Peptide Synthases / Q000032:analysis / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D014498:Uracil / Q000031:analogs & derivatives* / Q000032:analysis*; D014871:Water Microbiology
D000470:Alkaloids; D001427:Bacterial Toxins; D000087522:Cyanobacteria Toxins; D004269:DNA, Bacterial; D009097:Multienzyme Complexes; C089595:cylindrospermopsin; D014498:Uracil; D010453:Peptide Synthases