Four commercially available in vitro test systems (Candifast, E-test, Mycototal and spiral-gradient end point method), agar diffusion with 25-micrograms fluconazole paper test discs and 15-micrograms test tablets and agar dilution were compared with the microbroth dilution method for fluconazole susceptibility testing of 145 clinical isolates. In addition, the culture media provided or recommended by the manufacturers of the test systems were compared with high-resolution (HR) antifungal test medium. With all currently available culture media, growth problems (inhibition or delayed growth of the clinical isolates) occurred with solid or semisolid media. With minor improvements, HR medium demonstrated the most reproducible and comparable results (supplementation with asparagine and deletion of sodium hydrogen carbonate). The best correlation with microdilution was obtained by the agar dilution method (> 95% concordance) followed by the spiral-gradient end point method (85%), Candifast (83%), Mycototal (81%) and the E-test (78%). Regression analysis demonstrated good correlation between agar diffusion and micro-/agar dilution (r > 0.9).