Published May 9, 2020 | Version v1

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

  • 1. EA7361 'Structure, Dynamic, Function and Expression of Broad Spectrum b-Lactamases', University Paris-Saclay, LabEx Lermit, Faculty of Medicine, Le Kremlin- Bicêtre, France; Bacteriology-Hygiene Unit, Assistance Publique/Hôpitaux de Paris, Bicêtre Hospital, Le Kremlin- Bicêtre, France; Associated French National Reference Centre for Antibiotic Resistance: Carbapenemase-Producing Enterobacteriaceae, Le Kremlin-Bicêtre, France; Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France
  • 2. EA7361 'Structure, Dynamic, Function and Expression of Broad Spectrum b-Lactamases', University Paris-Saclay, LabEx Lermit, Faculty of Medicine, Le Kremlin- Bicêtre, France; Associated French National Reference Centre for Antibiotic Resistance: Carbapenemase-Producing Enterobacteriaceae, Le Kremlin-Bicêtre, France; Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France
  • 3. EA7361 'Structure, Dynamic, Function and Expression of Broad Spectrum b-Lactamases', University Paris-Saclay, LabEx Lermit, Faculty of Medicine, Le Kremlin- Bicêtre, France;
  • 4. EA7361 'Structure, Dynamic, Function and Expression of Broad Spectrum b-Lactamases', University Paris-Saclay, LabEx Lermit, Faculty of Medicine, Le Kremlin- Bicêtre, France; Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France
  • 5. Bacteriology-Hygiene Unit, Assistance Publique/Hôpitaux de Paris, Bicêtre Hospital, Le Kremlin- Bicêtre, France;
  • 6. Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France
  • 7. Evolution and Ecology of Resistance to Antibiotics Unit, Institut Pasteur—Assistance Publique/Hôpitaux de Paris—University Paris-Saclay, Paris, France; CNRS UMR3525, Paris, France

Description

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 with- in 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.

Notes

This work was partially funded by the University Paris-Sud, France. L.D., T.N. and R.A.B. are members of the Laboratory of Excellence in Research on Medication and Innovative Therapeutics (LERMIT) supported by a grant from the French National Research Agency (ANR-10-LABX-33), by the Joint Programming Initiative on Antimicrobial Resistance (JPIAMR) DesInMBL (ANR-14-JAMR-002) and by the European Union's Horizon 2020 Research and Innovation Program under Grant Agreement No. 773830 (Project MedVetKlebs, One Health EJP).

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Funding

European Commission
One Health EJP - Promoting One Health in Europe through joint actions on foodborne zoonoses, antimicrobial resistance and emerging microbiological hazards. 773830