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Related Experiment Videos

Optimizing testing of methicillin-resistant Staphylococcus species

C N Baker1, M B Huang, F C Tenover

  • 1Hospital Infections Program, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.

Diagnostic Microbiology and Infectious Disease
|July 1, 1994
PubMed
Summary
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Adding 2% NaCl to susceptibility testing media significantly improved accuracy for oxacillin resistance in Staphylococcus species. Higher concentrations (4-5% NaCl) led to unacceptable error rates across all tested methods.

Area of Science:

  • Microbiology
  • Clinical Diagnostics
  • Antimicrobial Resistance

Background:

  • Accurate oxacillin susceptibility testing for Staphylococcus species is crucial for guiding treatment.
  • Current methods lack standardized NaCl concentration recommendations for agar-based tests.
  • Inconsistent NaCl levels complicate accurate detection of oxacillin resistance.

Purpose of the Study:

  • To evaluate the impact of varying NaCl concentrations (0%, 2%, 4%, 5%) on oxacillin susceptibility testing of Staphylococcus species.
  • To determine the optimal NaCl concentration for agar dilution, Etest, and broth microdilution methods.
  • To compare test results with reference methods and mec gene detection.

Main Methods:

  • Tested 223 Staphylococcus strains (128 mec gene positive) using agar dilution, Etest, and broth microdilution.

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  • Varied NaCl concentration in Mueller-Hinton agar and broth from 0% to 5%.
  • Compared results against reference broth microdilution and mec gene hybridization assay.

    • Main Results:

    • Adding 2% NaCl increased Minimum Inhibitory Concentrations (MICs) for oxacillin-resistant strains (2-4 fold) without affecting susceptible strains.
    • Major and very major interpretive error rates decreased from 18.2-20.2% to <1% with 2% NaCl addition.
    • 4% and 5% NaCl concentrations resulted in >17% major error rates for all methods.

    Conclusions:

    • The addition of 2% NaCl to test media is essential for accurate oxacillin susceptibility testing of Staphylococcus species.
    • Standardizing 2% NaCl in agar dilution and Etest media improves diagnostic accuracy and reduces errors.
    • Higher NaCl concentrations (4-5%) should be avoided due to significant error induction.