Real-time quantitative LAMP (loop-mediated isothermal amplification of DNA) as a simple method for monitoring ammonia-oxidizing bacteria

Yoshiteru Aoi, Mariko Hosogai, Satoshi Tsuneda

    Research output: Contribution to journalArticle

    59 Citations (Scopus)

    Abstract

    Loop-mediated isothermal amplification (LAMP) of DNA is a novel technique for the amplification of DNA under isothermal conditions. For the first time, we applied this method to develop a simple and quantitative monitoring method for environmental microorganisms targeting amoA gene in ammonia-oxidizing bacteria. Quantitative analysis was performed first by measuring fluorescence derived from an intercalation dye using a real-time thermal cycler, and then by measuring the turbidity of the reaction solution using a real-time turbidimeter. As a result, it was possible to quantify the initial amoA DNA concentration from an environment with a sensitivity down to 102 DNA copies of target DNA and a dynamic range of 7-9 orders in magnitude. Background DNA from nontargeted bacteria (Pseudomonas denitrificans) that does not encode amoA gene did not affect the quantitative capability of LAMP. Over results suggested that the real-time LAMP is effective for monitoring microorganisms and their gene expression in environments.

    Original languageEnglish
    Pages (from-to)484-491
    Number of pages8
    JournalJournal of Biotechnology
    Volume125
    Issue number4
    DOIs
    Publication statusPublished - 2006 Oct 1

    Fingerprint

    Ammonia
    Amplification
    Bacteria
    DNA
    Monitoring
    Microorganisms
    Nucleic Acid Amplification Techniques
    Turbidimeters
    Genes
    Gene Targeting
    Environmental Monitoring
    Pseudomonas
    Turbidity
    Intercalation
    Gene expression
    Coloring Agents
    Hot Temperature
    Fluorescence
    Gene Expression
    Dyes

    Keywords

    • amoA gene
    • LAMP (loop-mediated isothermal amplification of DNA)
    • Quantification

    ASJC Scopus subject areas

    • Biotechnology

    Cite this

    Real-time quantitative LAMP (loop-mediated isothermal amplification of DNA) as a simple method for monitoring ammonia-oxidizing bacteria. / Aoi, Yoshiteru; Hosogai, Mariko; Tsuneda, Satoshi.

    In: Journal of Biotechnology, Vol. 125, No. 4, 01.10.2006, p. 484-491.

    Research output: Contribution to journalArticle

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