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    Important Factors in Adenosine Triphosphate Determinations Using Firefly Luciferase: Applicability of the Assay to Studies of Native Aquatic Bacteria

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    Accurate determinations of intracellular adenosine triphosphate (ATP) content in natural aquatic samples, using the firefly luciferase ATP assay, require attention to several important procedural factors. A rationale for peak height measurements is presented, along with a comparison of commercially available photometers. Purified luciferase, stabilized with dithiothreitol, is recommended for low-level ATP measurements. Inorganic acids were found to be the most efficient extracting agents for bacterial ATP. Limitations of the assay as a bacterial biomass measurement or enumeration technique are discussed, with the conclusion that the assay is most valuable in conjunction with other measures of total microbial deviations from a baseline.


    aquatic bacteria, firefly luciferase, adenosine triphosphate (ATP), bacterial biomass, ATP extraction, photometers, ATP flow system

    Author Information:

    Deming, JW
    Graduate assistant, University of Maryland, College Park, Md.

    Picciolo, GL
    Microbiologist, Research and Testing Staff, Bureau of Medical Devices, U.S. Food and Drug Administration, Silver Spring, Md.

    Chappelle, EW
    Physical scientist, Goddard Space Flight Center, Greenbelt, Md.

    Committee/Subcommittee: D19.24

    DOI: 10.1520/STP36007S