1887

Abstract

The mechanisms underlying colistin heteroresistance in are not fully understood. Here, we investigated the role of efflux in colistin-heteroresistant populations of a multidrug-resistant (MDR) clinical isolate.

Three colistin-resistant strain variants isolated from the same clinical sample were studied for the presence of heteroresistance to colistin by drug susceptibility testing, genotyping and drug resistance target mutation analysis. The existence of active efflux was studied by synergism assays with efflux inhibitors, real-time efflux activity measurements and analysis of the mRNA transcriptional levels of selected efflux pump genes in response to colistin.

All of the strain variants belong to the ST218, clonal complex 92, international clonal lineage II. Different colistin susceptibility levels were observed among the three strain variants, indicating that colistin-heteroresistant subpopulations were being selected upon exposure to colistin. No mutations were found in the genes and , which are associated with colistin resistance. The results showed the existence of synergistic interactions between efflux inhibitors and colistin and ethidium bromide. Real-time efflux assays demonstrated that the three strain variants had increased efflux activity that could be inhibited in the presence of the inhibitors. The efflux pump genes , , , , , and were found to be overexpressed in the strain variants in response to colistin exposure.

This study shows that efflux activity contributes to colistin heteroresistance in an MDR clinical isolate. The use of efflux inhibitors as adjuvants of the therapy can resensitize to colistin and prevent the emergence of drug resistance.

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2018-06-01
2020-01-21
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