Highly sensitive real-time PCR assay for quantification of toxic cyanobacteria based on microcystin synthetase a gene

Kazuhiro Furukawa, Naohiro Noda, Satoshi Tsuneda, Takeshi Saito, Tomoaki Itayama, Yuhei Inamori

    Research output: Contribution to journalArticle

    47 Citations (Scopus)

    Abstract

    The presence of cyanobacterial bloom in water supply reservoirs can cause potential health hazards. In this study, we aimed at the quantification of microcystin-producing cyanobacteria based on the microcystin synthetase A (mcyA) gene using real-time PCR. To perform a highly sensitive real-time PCR assay, the novel primer MSR-2R was designed and a coprecipitation DNA extraction method was used in this study. Cyanobacterial cells could be collected efficiently by coprecipitation with other bacteria suspended in solution even in the case of low concentrations of cyanobacteria. The detection limit of the method was found to be 8.8 cells per reaction. When cyanobacterial growth was monitored in pure culture, the cell concentration determined by real-time PCR positively correlated with the cell concentration determined from direct microscopic count. Furthermore, we could detect and quantify the mcyA gene in lake water samples using real-time PCR. It was concluded that the quantification of the mcyA gene based on real-time PCR is a powerful tool for the rapid quantification of microcystin-producing cyanobacteria in environmental samples.

    Original languageEnglish
    Pages (from-to)90-96
    Number of pages7
    JournalJournal of Bioscience and Bioengineering
    Volume102
    Issue number2
    DOIs
    Publication statusPublished - 2006 Aug

    Fingerprint

    Poisons
    Cyanobacteria
    Ligases
    Real-Time Polymerase Chain Reaction
    Assays
    Genes
    Coprecipitation
    Health hazards
    Water supply
    Lakes
    Bacteria
    DNA
    Water Supply
    Water
    Limit of Detection
    Cell Culture Techniques
    microcystin
    microcystin synthetase A
    Health
    Growth

    Keywords

    • cyanobacteria
    • mcyA gene
    • microcystin
    • Microcystis sp.
    • real-time PCR

    ASJC Scopus subject areas

    • Biotechnology
    • Bioengineering

    Cite this

    Highly sensitive real-time PCR assay for quantification of toxic cyanobacteria based on microcystin synthetase a gene. / Furukawa, Kazuhiro; Noda, Naohiro; Tsuneda, Satoshi; Saito, Takeshi; Itayama, Tomoaki; Inamori, Yuhei.

    In: Journal of Bioscience and Bioengineering, Vol. 102, No. 2, 08.2006, p. 90-96.

    Research output: Contribution to journalArticle

    Furukawa, Kazuhiro ; Noda, Naohiro ; Tsuneda, Satoshi ; Saito, Takeshi ; Itayama, Tomoaki ; Inamori, Yuhei. / Highly sensitive real-time PCR assay for quantification of toxic cyanobacteria based on microcystin synthetase a gene. In: Journal of Bioscience and Bioengineering. 2006 ; Vol. 102, No. 2. pp. 90-96.
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