Evaluation of susceptibility testing methods for Burkholderia cepacia complex: a comparison of broth microdilution,agar dilution,gradient strip and EUCAST disc diffusion

ObjectivesTo evaluate the accuracy and reproducibility of antimicrobial susceptibility testing methods in Burkholderia cepacia complex (BCC).MethodsMinocycline, ciprofloxacin, trimethoprim/sulphamethoxazole, meropenem, ceftazidime and chloramphenicol were tested against 155 BCC strains using broth microdilution at 35 ± 1°C (BMD₃₅) in triplicate, then BMD at 30 ± 1°C (BMD₃₀), agar dilution at 30°C and 35°C (AD₃₀ and AD₃₅), gradient strip (GS) and EUCAST standardized disc diffusion (DD) testing methods once.ResultsBMD₃₅ reproducibility ranged from 70% to 84.5% for all agents. Correlations of MICs from BMD₃₅ with BMD₃₀ ranged from 63% to 85%, with AD₃₅ from 32.9% to 87% and with GS methods from 36% to 83.9%. Essential agreement (EA) of MICs by GS with BMD₃₅ ranged from 62.6% (trimethoprim-sulphamethoxazole) to 83.9% (minocycline). EA of EUCAST DD zone diameters using CLSI breakpoint criteria was between 85.8% and 97.4%, however Very Major Errors (VME) for trimethoprim/sulphamethoxazole were 31%.ConclusionsBMD at 35 ± 1°C was poorly reproducible for most agents and no method showed acceptable performance. Of particular concern were the GS results. Although this is the most commonly used method for determining MICs in laboratories, there was poor correlation with BMD₃₅ for meropenem and trimethoprim/sulphamethoxazole. EUCAST DD correlated poorly with BMD₃₅ MICs. This study confirms that no susceptibility method is capable of providing reproducible and accurate MICs when testing BCC.