Question

Difficulty: HardInfectious Disease Mechanisms

A 32-year-old woman presents to the ambulatory care clinic with a painful, fluctuant soft tissue mass on her left thigh that has been enlarging over the past four days. She has no significant past medical history. Vital signs are within normal limits. An incision and drainage are performed, yielding purulent material that is sent for microbiologic analysis. Two days later, the culture grows *Staphylococcus aureus*. Antimicrobial susceptibility testing reveals resistance to oxacillin and erythromycin, but the isolate is initially reported as susceptible to clindamycin. The laboratory performs an additional disk diffusion assay by placing the erythromycin and clindamycin disks adjacent to each other on the agar plate. A distinct flattening of the clindamycin zone of inhibition adjacent to the erythromycin disk is observed. Based on these laboratory findings, the physician decides to prescribe an alternative oral antibiotic instead of clindamycin. Which of the following best describes the molecular mechanism responsible for the phenomenon observed in the adjacent disk diffusion testing?

  1. Post-transcriptional methylation of 23S ribosomal RNAAnswer
  2. B
    Active extrusion of the antimicrobial via ATP-dependent membrane efflux pumps
  3. C
    Acquisition of a chromosomal gene encoding an altered penicillin-binding protein
  4. D
    Enzymatic inactivation of the antimicrobial agent via plasmid-mediated nucleotidyltransferases
  5. E
    Point mutations in the quinolone resistance-determining region of DNA gyrase

Answer

Post-transcriptional methylation of 23S ribosomal RNA
The clinical vignette describes a positive D-test, which identifies inducible clindamycin resistance in staphylococci. When an isolate is resistant to erythromycin but appears susceptible to clindamycin in isolated testing, it may possess an *erm* (erythromycin ribosome methylation) gene. Erythromycin acts as an inducing agent, triggering the expression of this gene. The *erm* gene encodes an enzyme that methylates the 23S rRNA component of the 50S ribosomal subunit. This methylation alters the shared binding site for Macrolides, Lincosamides (clindamycin), and Streptogramin B (MLS_B resistance phenotype), rendering clindamycin ineffective in vivo despite initial in vitro susceptibility. Because of this, clindamycin should not be used for treatment, prompting the physician to choose an alternative.

Step-by-Step Solution

1
Analyze the susceptibility profile and the special laboratory test described in the vignette.
The isolate is MRSA (oxacillin resistant) that is erythromycin-resistant and clindamycin-susceptible, but shows a blunted clindamycin zone of inhibition when placed near erythromycin (a positive D-test).
Identifying the D-test is essential to determining the specific resistance phenotype being described.
2
Correlate the positive D-test with its underlying biological mechanism.
A positive D-test indicates inducible Macrolide-Lincosamide-Streptogramin B (MLS_B) resistance.
Erythromycin acts as an inducing agent that triggers the expression of a resistance gene (erm gene) in the bacteria.
3
Identify the molecular action of the erm gene product.
The erm gene encodes an enzyme that methylates the 23S rRNA of the 50S ribosomal subunit.
This target-site alteration prevents both macrolides (erythromycin) and lincosamides (clindamycin) from binding, explaining why clindamycin would fail in vivo despite appearing susceptible in standard isolated testing.

Key Concept

Inducible clindamycin resistance (MLS_B phenotype) in staphylococci is detected by a positive D-test and is mediated by the erm gene, which causes post-transcriptional methylation of the 23S rRNA binding site.
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