Get access

Identification of hepatotoxin-producing cyanobacteria by DNA-chip

Authors

  • Anne Rantala,

    1. Department of Applied Chemistry and Microbiology, University of Helsinki, Finland.
    Search for more papers by this author
  • Ermanno Rizzi,

    1. Institute of Biomedical Technologies, Italian National Research Council, Segrate (Mi), Italy.
    Search for more papers by this author
  • Bianca Castiglioni,

    1. Institute of Biomedical Technologies, Italian National Research Council, Segrate (Mi), Italy.
    Search for more papers by this author
    • Present address: Institute of Agricultural Biology and Biotechnology, Italian National Research Council, Milan, Italy.

  • Gianluca De Bellis,

    1. Institute of Biomedical Technologies, Italian National Research Council, Segrate (Mi), Italy.
    Search for more papers by this author
  • Kaarina Sivonen

    Corresponding author
    1. Department of Applied Chemistry and Microbiology, University of Helsinki, Finland.
      *E-mail kaarina.sivonen@helsinki.fi; Tel. (+358) 919159270; Fax (+358) 9 19159322.
    Search for more papers by this author

*E-mail kaarina.sivonen@helsinki.fi; Tel. (+358) 919159270; Fax (+358) 9 19159322.

Summary

We developed a new tool to detect and identify hepatotoxin-producing cyanobacteria of the genera Anabaena, Microcystis, Planktothrix, Nostoc and Nodularia. Genus-specific probe pairs were designed for the detection of the microcystin (mcyE) and nodularin synthetase genes (ndaF) of these five genera to be used with a DNA-chip. The method couples a ligation detection reaction, in which the polymerase chain reaction (PCR)-amplified mcyE/ndaF genes are recognized by the probe pairs, with a hybridization on a universal microarray. All the probe pairs specifically detected the corresponding mcyE/ndaF gene sequences when DNA from the microcystin- or nodularin-producing cyanobacterial strains were used as template in the PCR. Furthermore, the strict specificity of detection enabled identification of the potential hepatotoxin producers. Detection of the genes was very sensitive; only 1–5 fmol of the PCR product were needed to produce signal intensities that exceeded the set background threshold level. The genus-specific probe pairs also reliably detected potential microcystin producers in DNA extracted from six lake and four brackish water samples. In lake samples, the same microcystin producers were identified with quantitative real-time PCR analysis. The specificity, sensitivity and ability of the DNA-chip in simultaneously detecting all the main hepatotoxin producers make this method suitable for high-throughput analysis and monitoring of environmental samples.

Get access to the full text of this article

Ancillary