Concomitant carriage of KPC-producing and non-KPC-producing Klebsiella pneumoniae ST512 within a single patient

Author:

Jousset Agnès B1234,Bonnin Rémy A134ORCID,Takissian Julie1,Girlich Delphine14,Mihaila Liliana2,Cabanel Nicolas4,Dortet Laurent1234,Glaser Philippe45,Naas Thierry1234ORCID

Affiliation:

1. EA7361 ‘Structure, Dynamic, Function and Expression of Broad Spectrum β-Lactamases’, University Paris-Saclay, LabEx Lermit, Faculty of Medicine, Le Kremlin-Bicêtre, France

2. Bacteriology-Hygiene Unit, Assistance Publique/Hôpitaux de Paris, Bicêtre Hospital, Le Kremlin-Bicêtre, France

3. Associated French National Reference Centre for Antibiotic Resistance: Carbapenemase-Producing Enterobacteriaceae, Le Kremlin-Bicêtre, France

4. Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France

5. CNRS UMR3525, Paris, France

Abstract

Abstract Background KPC-producing Klebsiella pneumoniae of clonal group 258 are prominent in healthcare settings in many regions of the world. The blaKPC gene is mostly carried by a multireplicon IncFIIk-IncFI plasmid suspected to be highly compatible and stable in this genetic background. Here, we analysed the genetic diversity of an ST512 K. pneumoniae population in a single patient. Methods Twelve K. pneumoniae isolates (n = 5 from urine samples and n = 7 from rectal swabs) were recovered from one patient over a 2 month period. Antimicrobial susceptibility testing, plasmid extraction and WGS were performed on all isolates. The first K. pneumoniae isolate, D1, was used as a reference for phylogenetic analysis. Results Antimicrobial susceptibility testing, plasmid analysis and WGS revealed concomitant carriage of carbapenem-resistant and carbapenem-susceptible K. pneumoniae isolates of ST512, with the absence of the entire blaKPC-carrying plasmid in the susceptible population. Furthermore, 14 other genetic events occurred within the genome, including 3 chromosomal deletions (of 71 kb, 33 kb and 11 bp), 2 different insertions of ISKpn26 and 9 SNPs. Interestingly, most of the events occurred in the same chromosomal region that has been deleted independently several times, probably after homologous recombination involving 259 bp repeated sequences. Conclusions Our study revealed (to the best of our knowledge) the first case of in vivo blaKPC-carrying plasmid curing and a wide within-patient genetic diversity of a single K. pneumoniae ST512 clone over a short period of carriage. This within-patient diversity must be taken into account when characterizing transmission chains using WGS during nosocomial outbreaks.

Funder

University Paris-Sud

Laboratory of Excellence in Research on Medication and Innovative Therapeutics

French National Research Agency

Joint Programming Initiative on Antimicrobial Resistance

European Union’s Horizon 2020 Research and Innovation Program

Publisher

Oxford University Press (OUP)

Subject

Infectious Diseases,Pharmacology (medical),Pharmacology,Microbiology (medical)

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