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Clin Microbiol Infect. 2014 Apr;20(4):O255-66. doi: 10.1111/1469-0691.12373. Epub 2013 Oct 17.

Development of the EUCAST disk diffusion antimicrobial susceptibility testing method and its implementation in routine microbiology laboratories.

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EUCAST Laboratory for Antimicrobial Susceptibility Testing, Växjö, Sweden.


With the support of ESCMID and European countries, EUCAST has developed a disk diffusion test with zone diameter breakpoints correlated with the EUCAST clinical MIC breakpoints. The development of the EUCAST disk diffusion method and quality control criteria are described, together with guidance on quality control and implementation of the method in clinical microbiology laboratories. The method includes the use of Mueller-Hinton agar without supplements for non-fastidious organisms and with 5% mechanically defibrinated horse blood and 20 mg/L β-NAD for fastidious organisms, a standardized inoculum resulting in confluent growth, an incubation time of 16-20 h, a reading guide on how to read zone diameters on individual species-agent combinations and zone diameter breakpoints calibrated to the EUCAST clinical MIC breakpoints. EUCAST recommendations are described in detail and updated regularly on the EUCAST website (


Antimicrobial susceptibility testing; European Committee on Antimicrobial Susceptibility Testing; MIC; Mueller-Hinton agar; disk diffusion; zone diameter breakpoints

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