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Bacterial characterization by flow cytometry.

M A Van Dilla, R G Langlois, D Pinkel

    Science (New York, N.Y.)
    |May 6, 1983
    PubMed
    Summary
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    This study introduces a novel flow cytometry method for analyzing bacterial DNA base composition and content. The technique accurately characterizes bacteria in urine samples, aiding urinary tract infection diagnosis.

    Area of Science:

    • Microbiology
    • Molecular Biology
    • Biotechnology

    Background:

    • Accurate characterization of bacterial DNA is crucial for understanding microbial populations and diagnosing infections.
    • Traditional methods may face challenges with complex sample matrices like urine, leading to interference from host cells or debris.

    Purpose of the Study:

    • To develop and validate a dual-beam flow cytometry method for analyzing bacterial DNA content and base composition.
    • To assess the utility of this method for characterizing bacteria in clinical samples, specifically urine from patients with urinary tract infections.

    Main Methods:

    • Bacteria were analyzed using a dual-beam flow cytometer.
    • Double staining with chromomycin A3 (binds GC-rich DNA) and Hoechst 33258 (binds AT-rich DNA) was employed.

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  • Measurements included cellular DNA content, base composition, cell concentration, and proliferative state.

    • Main Results:

    • The ratio of chromomycin A3 to Hoechst 33258 signal correlated with the guanine-cytosine content of bacterial DNA across six cultured species.
    • The method successfully characterized bacteria in urine samples from patients with urinary tract infections.
    • Analysis was performed without interference from host cell DNA, debris, or other particulates.

    Conclusions:

    • This novel flow cytometry technique provides a reliable method for assessing bacterial DNA content and base composition.
    • The method offers a robust approach for analyzing bacteria in complex clinical samples, improving diagnostic capabilities for urinary tract infections.