Multiplex PCR assay for Cylindrospermopsis raciborskii and cylindrospermopsin-producing cyanobacteria

Authors

  • Kim M. Fergusson,

    1. Cooperative Research Centre for Water Quality and Treatment, Australian Water Quality Centre, SA Water Corporation, Private Mail Bag 3, Salisbury, South Australia 5108, Australia, and School of Pharmaceutical, Molecular and Biomedical Sciences, University of South Australia, Mawson Lakes Campus, Mawson Lakes, South Australia 5095, Australia
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  • Christopher P. Saint

    Corresponding author
    1. Cooperative Research Centre for Water Quality and Treatment, Australian Water Quality Centre, SA Water Corporation, Private Mail Bag 3, Salisbury, South Australia 5108, Australia, and School of Pharmaceutical, Molecular and Biomedical Sciences, University of South Australia, Mawson Lakes Campus, Mawson Lakes, South Australia 5095, Australia
    • Australian Water Quality Centre, SA Water Corporation, Private Mail Bag 3, Salisbury, South Australia 5108, Australia
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Abstract

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. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 120–125, 2003.

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