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Related Experiment Videos

Rhinovirus plaque formation in WI-38 cells with methylcellulose overlay.

T M Dolan, J D Fenters, P A Fordyce

    Applied Microbiology
    |September 1, 1968
    PubMed
    Summary
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    A new plaque assay system using methylcellulose overlay reliably detects five rhinoviruses. This method is sensitive to temperature and pH, offering a robust tool for rhinovirus research.

    Area of Science:

    • Virology
    • Cell Biology

    Background:

    • Rhinoviruses are a common cause of human respiratory infections.
    • Sensitive and reliable plaque assay systems are crucial for studying rhinovirus replication and characterization.

    Purpose of the Study:

    • To describe a sensitive and reliable plaque assay system for quantifying five specific rhinoviruses.
    • To evaluate the impact of various conditions on rhinovirus plaque formation.

    Main Methods:

    • Utilized freshly prepared methylcellulose overlay and human embryonic diploid cells for plaque assays.
    • Incubated cultures for 6-14 days to observe plaque formation.
    • Tested the effects of pH, temperature, and magnesium ions on plaque characteristics.

    Main Results:

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    • Rhinoviruses 1A, 2, 6, and 13 formed circular plaques (2 mm) in 6-7 days; Rhinovirus 17 formed feather plaques (1-2 mm) in 14 days.
    • Plaque counts for rhinoviruses 1A and 13 were stable across pH 6.9-7.5.
    • Plaque size and PFU values for high passage rhinoviruses 1A and 13 were consistent at 26°C, 31°C, and 36°C.
    • Rhinoviruses showed sensitivity to 40°C incubation and 50°C heating.
    • Magnesium ions enhanced plaques in agar overlays but not in methylcellulose overlays.

    Conclusions:

    • The described methylcellulose overlay plaque assay is a sensitive and reliable method for quantifying specific rhinoviruses.
    • The assay is robust to variations in pH and temperature within tested ranges.
    • Understanding plaque formation characteristics aids in rhinovirus identification and study.