Comparison of disc diffusion assay with the CLSI reference method (M27-A2) for testing in vitro posaconazole activity against common and uncommon yeasts

J Antimicrob Chemother. 2008 Jan;61(1):135-8. doi: 10.1093/jac/dkm442. Epub 2007 Nov 22.

Abstract

Objectives: To evaluate the suitability of disc diffusion (DD) assay for testing posaconazole activity and to corroborate its activity against recently isolated yeasts by the CLSI reference microdilution M27-A2 method.

Methods: A total of 224 yeast isolates (7 species with 52 to 11 isolates each, and 15 species with 1 to 6 isolates) were evaluated, 125 were recent bloodstream isolates, 30 isolates from other sources and six ATCC isolates that included amphotericin B-resistant Candida albicans ATCC 200955, Candida lusitaniae (ATCC 200950, 200951, 200952 and 200953) and amphotericin B- and itraconazole-resistant Candida tropicalis ATCC 200956. MICs were determined at 24 and 48 h by following the CLSI guidelines, document M27-A2. DD testing was performed by following CLSI M44-A document with 5 microg posaconazole discs. Inhibition zone diameters were measured at the transition point at which growth decreased at both 24 and 48 h.

Results: DD showed very good reproducibility, with coefficient of variability median value 4.56. Posaconazole demonstrated good in vitro activity against all clinical isolates, including the emerging species and amphotericin B-resistant ATCC isolates except for C. tropicalis ATCC 200956 (posaconazole MIC >or= 16 mg/L). Only 1.5% and 4.1% of isolates were inhibited by >2 mg/L posaconazole at 24 and 48 h. Good correlation was obtained between methods (R = 0.763 at 24 h and 0.602 at 48 h). DD detected posaconazole-resistant isolates (MIC > 2 mg/L).

Conclusions: DD could be an alternative to the microdilution reference method, as no major discrepancies were detected.

Publication types

  • Comparative Study

MeSH terms

  • Antifungal Agents / pharmacology*
  • Candida / drug effects*
  • Candida / isolation & purification
  • Candidiasis / microbiology
  • Drug Resistance, Fungal / drug effects*
  • Humans
  • Microbial Sensitivity Tests / methods
  • Reference Standards
  • Reproducibility of Results
  • Triazoles / pharmacology*

Substances

  • Antifungal Agents
  • Triazoles
  • posaconazole