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Related Experiment Videos

Plaque formation by virulent Shigella flexneri.

E V Oaks, M E Wingfield, S B Formal

    Infection and Immunity
    |April 1, 1985
    PubMed
    Summary

    A novel plaque assay effectively models virulent shigellae

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    Area of Science:

    • Microbiology
    • Cell Biology
    • Infectious Diseases

    Background:

    • Shigella species are invasive bacterial pathogens causing bacillary dysentery.
    • Understanding Shigella's intracellular replication and spread is crucial for developing interventions.

    Purpose of the Study:

    • To develop and validate an in vitro plaque assay for studying virulent shigellae.

    Main Methods:

    • HeLa cell monolayers were cultured and overlaid with agarose containing gentamicin to prevent extracellular growth.
    • Plaque formation was assessed under varying temperature conditions and in the presence of specific inhibitors.
    • Virulence factors of Shigella isolated from plaques were characterized.

    Main Results:

    • The assay successfully visualized plaques characterized by host cell lysis and bacterial spread.
    • Plaque formation was dependent on temperature (37°C) and inhibited by chloramphenicol and nalidixic acid.
    • All tested virulent Shigella serotypes formed plaques, which were associated with gentamicin sensitivity and a 140-MDa plasmid.

    Conclusions:

    • The developed plaque assay is a reliable tool for investigating Shigella's intracellular and intercellular pathogenic mechanisms.
    • This assay differentiates virulent, invasive Shigella strains from noninvasive ones.
    • The findings highlight the importance of temperature and specific genetic elements in Shigella pathogenesis.

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