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Estimation of Mycoplasma pneumoniae inoculum size by rate of tetrazolium reduction

W Bredt

    Journal of Clinical Microbiology
    |July 1, 1976
    PubMed

    Insights

    The reduction of 2,3,5-triphenyltetrazolium chloride by Mycoplasma pneumoniae, indicated by red formazan, is directly related to the inoculum size. This new method allows for the estimation of Mycoplasma pneumoniae cells when traditional dilution counts are not feasible.

    Area of Science:

    • Microbiology
    • Cellular Metabolism

    Background:

    • Mycoplasma pneumoniae is a significant human pathogen.
    • Accurate quantification of M. pneumoniae is crucial for diagnosis and research.
    • Traditional counting methods like dilution plating can be challenging for certain M. pneumoniae preparations.

    Purpose of the Study:

    • To develop a novel method for quantifying Mycoplasma pneumoniae.
    • To establish a colorimetric assay based on tetrazolium salt reduction.

    Main Methods:

    • Utilized 2,3,5-triphenyltetrazolium chloride (TTC) as a substrate.
    • Measured the rate of formazan production, a byproduct of TTC reduction by viable M. pneumoniae cells.
    • Correlated formazan appearance with bacterial inoculum size.

    Main Results:

    • The rate of formazan formation was directly dependent on the initial inoculum size of M. pneumoniae.
    • The colorimetric assay provided a quantitative measure of M. pneumoniae presence.
    • This method proved effective for estimating cell numbers in preparations unsuitable for dilution methods.

    Conclusions:

    • The tetrazolium salt reduction assay is a viable method for quantifying Mycoplasma pneumoniae.
    • This technique offers an alternative for estimating M. pneumoniae in challenging samples.
    • The assay's sensitivity to inoculum size allows for reliable cell quantification.

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