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A sensitive assay for Staphylococcus aureus nucleases.

J K Kohli, B V Vakil, M S Patil

    Indian Journal of Biochemistry & Biophysics
    |October 1, 1989
    PubMed
    Summary
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    A new assay accurately detects staphylococcal nuclease using radiolabeled DNA. This method is highly sensitive for identifying Staphylococcus aureus contamination in food and water, surpassing traditional techniques.

    Area of Science:

    • Microbiology
    • Biochemistry
    • Analytical Chemistry

    Background:

    • Staphylococcal nuclease is an important enzyme produced by Staphylococcus aureus.
    • Existing methods for detecting staphylococcal nuclease and S. aureus contamination are often insensitive.
    • There is a need for a more sensitive assay to quantify staphylococcal nuclease levels and microbial contamination.

    Purpose of the Study:

    • To develop a highly sensitive assay for staphylococcal nuclease.
    • To enable accurate comparison of nuclease production among different Staphylococcus aureus strains.
    • To detect low levels of Staphylococcus aureus contamination in food and water samples.

    Main Methods:

    • Incubation of enzyme samples with heat-denatured, [3H]thymidine-labeled DNA from E. coli.

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  • Precipitation of undigested DNA using trichloroacetic acid.
  • Measurement of radioactivity in the acid-soluble supernatant, representing released nucleotides.

    • Main Results:

    • The developed assay demonstrates high sensitivity for staphylococcal nuclease detection.
    • The assay effectively differentiates nuclease levels produced by various Staphylococcus aureus strains.
    • It successfully identifies Staphylococcus aureus contamination at levels undetectable by conventional methods.

    Conclusions:

    • A novel, sensitive radiochemical assay for staphylococcal nuclease has been established.
    • This assay provides a valuable tool for microbial strain comparison and contamination assessment.
    • The method significantly improves upon the limitations of existing spectrophotometric and toluidine blue-DNA assays.