Abstract

ABSTRACTUp to 4-fold differences in zinc concentrations have been observed in commercial broth routinely utilized for susceptibility testing via manual broth microdilution. Herein, we report the concentration of zinc in the broth of common automated susceptibility testing (AST) platforms (Vitek, MicroScan, BD Phoenix, and Sensititre). For AST platforms with lyophilized broth contents (Vitek and MicroScan), wells were rehydrated with appropriate diluent, and contents were aliquoted out for zinc assay. Aliquots from the manufacturer-specific broth (premade cation-adjusted Mueller-Hinton broth [caMHB]) for BD Phoenix and Sensititre were also assayed by inductively coupled plasma mass spectrometry. Up to a 10-fold difference in zinc concentrations was observed across the 4 platforms (MicroScan: 0.46 mg/L; BD Phoenix: 1.16 mg/L; Vitek: 1.22 mg/L; Sensititre: 4.49 mg/L). Attention should be given to the supraphysiologic and variable zinc concentrations observed in broth used in automated platforms and the subsequent implications for susceptibility testing of metallo-β-lactamase (MBL)-harboring isolates. This variability also hampers efforts to develop a standardized method to uniformly reduce zinc concentrations in broth and mimic physiologic zinc conditions.IMPORTANCE Growing data on the impact of extracellular zinc concentration on metallo-β-lactamase-mediated resistance has shed light on the importance of susceptibility testing media. However, there are no studies documenting the amount of zinc in commonly utilized automated susceptibility testing (AST) platforms. This study reveals supraphysiologic zinc concentrations as well as large zinc variability among AST platforms and highlights the challenges this raises in the development of zinc-limited media.

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