Molecular Characterization of Tigecycline Non-Susceptibility among Extensively Drug-Resistant Acinetobacter baumannii Isolates of Clinical Origin

Abstract

<b><i>Introduction:</i></b> Tigecycline (TGC) is one of the last-resort therapeutic agents for treating infections caused by extensively drug resistant <i>Acinetobacter baumannii</i> isolates. Although resistance to TGC is not common, non-susceptible <i>A. baumannii</i> (NSAB) isolates have been described. In the current study, we aimed to assess the molecular mechanisms mediating TGC non-susceptibility in 5 clinical isolates of <i>A. baumannii</i> with reduced susceptibility to TGC. <b><i>Methods:</i></b> Susceptibility of isolates to TGC as well as various classes of antibiotics was evaluated by broth dilution and disk diffusion methods, respectively. The presence of <i>tetX</i> and <i>tetX1</i> genes was investigated by PCR. The nucleotide sequences of <i>adeR</i> and <i>adeS</i> genes were assessed by PCR amplicon sequencing. To evaluate the association between reduced susceptibility to TGC and upregulation of AdeABC efflux pump, transcriptional level of <i>adeB</i> gene was quantified by RT-qPCR analysis. <b><i>Results:</i></b> All 5 TGC-NSAB isolates had a TGC MIC of ≥4 mg/L and were resistant to all antimicrobials tested by disk diffusion method except for minocycline and doxycycline for which a susceptibility rate of 40% and 20% was observed, respectively. The <i>tetX/X1</i> genes were not detected in any isolates. All TGC non-susceptible isolates harbored genetic alterations in the <i>adeRS</i> operon, including AdeS G186V, N268H, and D60N and AdeR A136V and V120I substitutions among, which G186V and D60N were predicted by PROVEAN tool analysis as inactivating alterations. Reduced TGC susceptibility was associated with upregulation of AdeABC efflux pump in all TGC non-susceptible isolates. <b><i>Conclusion:</i></b> It can be concluded from our results that reduced susceptibility to TGC in the studied isolates was mainly mediated by genetic alterations in the AdeRS system, which resulted in overexpression of AdeABC efflux pump. Emergence of TGC non-susceptibility among isolates that had not been previously exposed to TGC is an issue of great concern.