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Quantitative, radial diffusion slide assay for staphylocoagulase

J D Kohl, M G Johnson

    Applied and Environmental Microbiology
    |February 1, 1980
    PubMed
    Summary
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    A new radial diffusion assay quantifies staphylocoagulase in Staphylococcus aureus cultures. This simple method requires minimal sample volume and no enzyme purification, enabling objective comparison of coagulase production.

    Area of Science:

    • Microbiology
    • Biochemistry
    • Diagnostic Assays

    Background:

    • Staphylocoagulase is a key virulence factor produced by Staphylococcus aureus.
    • Accurate quantification of staphylocoagulase is important for understanding bacterial pathogenesis and for diagnostic purposes.
    • Existing methods for coagulase detection may require complex procedures or large sample volumes.

    Purpose of the Study:

    • To develop a simple, quantitative radial diffusion assay for staphylocoagulase.
    • To enable objective comparison of coagulase production in different Staphylococcus aureus cultures.
    • To establish a method requiring minimal sample volume and no prior enzyme purification.

    Main Methods:

    • Development of a radial diffusion assay using microscope slides coated with agar containing plasma and fibrinogen.

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  • Application of the assay to culture fluids of Staphylococcus aureus.
  • Quantitative analysis of zones of precipitation to determine coagulase activity.

    • Main Results:

    • A simple and quantitative radial diffusion assay for staphylocoagulase was successfully developed.
    • The assay requires only small sample volumes (7 microliters) per test.
    • No prior purification of staphylocoagulase from culture fluids was necessary.

    Conclusions:

    • The developed radial diffusion assay is a suitable method for objectively comparing the relative amounts of staphylocoagulase produced by different Staphylococcus aureus cultures.
    • This assay offers a simplified and efficient approach for staphylocoagulase quantification.
    • The method's ease of use and minimal sample requirement make it valuable for routine laboratory applications.