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The prevalence of azithromycin resistance in Neisseria gonorrhoeae is increasing in numerous populations worldwide. To characterize the genetic pathways leading to high-level azithromycin resistance. A customized morbidostat was used to subject two N. gonorrhoeae reference strains (WHO-F and WHO-X) to dynamically sustained azithromycin pressure. We tracked stepwise evolution of resistance by whole genome sequencing. Within 26 days, all cultures evolved high-level azithromycin resistance. Typically, the first step towards resistance was fo
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