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Related Experiment Videos

MBCs for Staphylococcus aureus as determined by macrodilution and microdilution techniques.

C J Shanholtzer, L R Peterson, M L Mohn

    Antimicrobial Agents and Chemotherapy
    |August 1, 1984
    PubMed
    Summary

    Reproducible Minimum Bactericidal Concentration (MBC) testing of Staphylococcus aureus requires careful methodology. Standard macrodilution and multiprong inoculation methods proved unreliable, with optimal results achieved using a 100-microliter pipette for subculturing microdilution panels.

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    Area of Science:

    • Microbiology
    • Antimicrobial Susceptibility Testing

    Background:

    • Accurate Minimum Bactericidal Concentration (MBC) determination is crucial for guiding antibiotic therapy against Staphylococcus aureus.
    • Standardized and reproducible methods for MBC testing are essential for reliable clinical and research applications.

    Purpose of the Study:

    • To evaluate the reproducibility of standard macrodilution tubes and commercial microdilution trays for MBC testing of Staphylococcus aureus.
    • To compare different subculturing techniques for microdilution panels to identify the most reliable method.

    Main Methods:

    • Tested clindamycin, methicillin, cephalothin, gentamicin, and vancomycin against 67 Staphylococcus aureus clinical isolates.
    • Compared standard macrodilution with commercial microdilution trays.
    • Evaluated various subculturing methods including multiprong inoculators, 1-microliter loops, 10-microliter pipettes, and 100-microliter pipettes.

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    Main Results:

    • Standard macrodilution failed to yield reproducible MBC results, even with strict protocols.
    • Incubation conditions (shaking vs. stationary) and vortexing techniques significantly impacted bacterial regrowth.
    • Subculturing commercial microdilution panels using a 100-microliter pipette after stirring provided the most reproducible MBC results.

    Conclusions:

    • Standard macrodilution methods are not reliable for MBC testing of Staphylococcus aureus.
    • Careful optimization of incubation and subculturing techniques is critical for reproducible MBC testing.
    • Aspirating the entire 100-microliter volume from microdilution wells and spreading onto agar offers the highest reproducibility.