EUCAST disc diffusion criteria for the detection of mecA-Mediated β-lactam resistance in Staphylococcus pseudintermedius: oxacillin versus cefoxitin

Clin Microbiol Infect. 2020 Jan;26(1):122.e1-122.e6. doi: 10.1016/j.cmi.2019.05.002. Epub 2019 May 17.

Abstract

Objectives: Until recently, the European Committee on Antimicrobial Susceptibility Testing (EUCAST) recommended the cefoxitin disc to screen for mecA-mediated β-lactam resistance in Staphylococcus pseudintermedius. A recent study indicated that cefoxitin was inferior to oxacillin in this respect. We have re-evaluated cefoxitin and oxacillin discs for screening for methicillin resistance in S. pseudintermedius.

Methods: We included 224 animal and human S. pseudintermedius isolates from Europe (n = 108) and North America (n = 116), of which 109 were mecA-positive. Disc diffusion was performed per EUCAST recommendations using 30-μg cefoxitin and 1-μg oxacillin discs from three manufacturers and Mueller-Hinton agar from two manufacturers.

Results: Cefoxitin inhibition zones ranged from 6 to 33 mm for mecA-positive S. pseudintermedius (MRSP) and from 29 to 41 mm for mecA-negative S. pseudintermedius (MSSP). The corresponding oxacillin zone intervals were 6-20 mm and 19-30 mm. For cefoxitin 16% (95% CI 14.8-18.0%) of the isolates were in the area where positive and negative results overlapped. For oxacillin the corresponding number was 2% (1.6-2.9%). For oxacillin a breakpoint of susceptible (S) ≥ 20 mm and resistant (R) <20 mm resulted in only 0.4% and 1.1% very major error and major error rates respectively.

Conclusions: This investigation confirms that the 1-μg oxacillin disc predicts mecA-mediated methicillin resistance in S. pseudintermedius better than the 30-μg cefoxitin disc. For a 1-μg oxacillin disc we propose that 20 mm should be used as cut off for resistance, i.e. isolates with a zone diameter <20 mm are resistant to all β-lactam antibiotics except those with activity against methicillin-resistant staphylococci.

Keywords: Cefoxitin; Methicillin resistance; Oxacillin; Staphylococcus pseudintermedius; Susceptibility testing; mecA-positive S.pseudintermedius.

MeSH terms

  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Cefoxitin / pharmacology*
  • Disk Diffusion Antimicrobial Tests / methods*
  • Disk Diffusion Antimicrobial Tests / standards
  • Humans
  • Microbial Sensitivity Tests
  • Oxacillin / pharmacology*
  • Staphylococcal Infections / microbiology
  • Staphylococcus / drug effects*
  • Staphylococcus / enzymology
  • beta-Lactam Resistance*

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • MecA protein, Staphylococcus pseudintermedius
  • Cefoxitin
  • Oxacillin

Supplementary concepts

  • Staphylococcus pseudintermedius