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Related Experiment Video

Updated: Mar 28, 2026

Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds
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Optimized In Vitro Antibiotic Susceptibility Testing Method for Small-Colony Variant Staphylococcus aureus.

Mimi R Precit1, Daniel J Wolter2, Adam Griffith3

  • 1University of Washington, Seattle, Washington, USA.

Antimicrobial Agents and Chemotherapy
|January 6, 2016
PubMed
Summary

Standardized antibiotic susceptibility testing (AST) for Staphylococcus aureus small-colony variants (SCVs) is challenging. This study identifies optimal media and methods for SCV AST, aiding in chronic infection treatment.

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Area of Science:

  • Microbiology
  • Infectious Diseases
  • Antimicrobial Resistance

Background:

  • Staphylococcus aureus small-colony variants (SCVs) are linked to persistent infections and poor outcomes.
  • Standard antibiotic susceptibility testing (AST) methods are unreliable for SCVs due to poor growth and reversion.
  • Predicting SCV antibiotic resistance based on clinical history or related normal-colony (NC) isolates is difficult.

Purpose of the Study:

  • To establish reproducible methods for AST of S. aureus SCVs.
  • To determine if SCV susceptibilities can be predicted by auxotrophy, treatment history, or isogenic NC isolate data.

Main Methods:

  • Evaluated SCV growth and reversion on 11 agar media, selecting two optimal media.
  • Performed disk diffusion and Etest AST on 86 SCVs and 28 isogenic NCs.
  • Analyzed SCV auxotrophy, NC antibiograms, and patient treatment history for predictive value.

Main Results:

  • Brain heart infusion and Mueller-Hinton agars supplemented with hemin, menadione, and thymidine supported optimal SCV growth and stability.
  • SCVs showed predictable nonsusceptibility to trimethoprim-sulfamethoxazole or aminoglycosides based on auxotrophy.
  • Varied nonsusceptibility was observed for fluoroquinolones, macrolides, and others; mecA-positive SCVs were cefoxitin-resistant.
  • All isolates remained susceptible to quinupristin-dalfopristin, vancomycin, minocycline, linezolid, chloramphenicol, and tigecycline.

Conclusions:

  • Developed a reliable AST method for S. aureus SCVs using supplemented BHI and MH agars.
  • SCV auxotrophy predicts resistance to certain antibiotics, but other resistance patterns are variable.
  • Predictive value of treatment history and isogenic NC isolates for SCV susceptibility is limited.
  • This validated AST method and findings can guide treatment decisions for challenging SCV infections.