title: IncFIIk plasmid harbouring an amplification of 16S rRNA methyltransferase-encoding gene rmtH associated with mobile element ISCR2.
abstract: To investigate the resistance mechanisms and genetic support underlying the high resistance level of the Klebsiella pneumoniae strain CMUL78 to aminoglycoside and β-lactam antibiotics.
  
  
  
  Antibiotic susceptibility was assessed by the disc diffusion method and MICs were determined by the microdilution method. Antibiotic resistance genes and their genetic environment were characterized by PCR and Sanger sequencing. Plasmid contents were analysed in the clinical strain and transconjugants obtained by mating-out assays. Complete plasmid sequencing was performed with PacBio and Illumina technology.
  
  
  
  Strain CMUL78 co-produced the 16S rRNA methyltransferase (RMTase) RmtH, carbapenemase OXA-48 and ESBL SHV-12. The rmtH- and blaSHV-12-encoding genes were harboured by a novel ∼115 kb IncFIIkplasmid designated pRmtH, and blaOXA-48by a ∼62 kb IncL/M plasmid related to pOXA-48a. pRmtH plasmid possessed seven different stability modules, one of which is a novel hybrid toxin-antitoxin system. Interestingly, pRmtH plasmid harboured a 4-fold amplification of an rmtH-ISCR2 unit arranged in tandem and inserted within a novel IS26-based composite transposon designated Tn6329.
  
  
  
  This is the first known report of the 16S RMTase-encoding gene rmtH in a plasmid. The rmtH-ISCR2 unit was inserted in a composite transposon as a 4-fold tandem repeat, a scarcely reported organization.
authors: CHU Clermont-Ferrand, Laboratoire de Bactériologie Clinique, Clermont-Ferrand, France.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.|Clermont Université, Université d'Auvergne, M2iSH, Clermont-Ferrand, France.|UMR INSERM 1071, Clermont-Ferrand, France.|USC INRA2018, Clermont-Ferrand, France.
authors: Racha
authors: R
authors: Beyrouthy
authors: CHU Clermont-Ferrand, Laboratoire de Bactériologie Clinique, Clermont-Ferrand, France.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.|Clermont Université, Université d'Auvergne, M2iSH, Clermont-Ferrand, France.|UMR INSERM 1071, Clermont-Ferrand, France.|USC INRA2018, Clermont-Ferrand, France.
authors: Frederic
authors: F
authors: Robin
authors: Laboratoire Microbiologie Santé et Environnement (LMSE), Ecole Doctorale en Sciences et Technologies et Faculté de Santé Publique, Université Libanaise, Tripoli, Lebanon.
authors: Monzer
authors: M
authors: Hamze
authors: CHU Clermont-Ferrand, Laboratoire de Bactériologie Clinique, Clermont-Ferrand, France rbonnet@chu-clermontferrand.fr.|Centre National de Référence de la Résistance aux Antibiotiques, laboratoire associé, Clermont-Ferrand, France.|Clermont Université, Université d'Auvergne, M2iSH, Clermont-Ferrand, France.|UMR INSERM 1071, Clermont-Ferrand, France.|USC INRA2018, Clermont-Ferrand, France.
authors: Richard
authors: R
authors: Bonnet
mesh_terms: D000617:Aminoglycosides / Q000494:pharmacology*; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D022761:Gene Transfer, Horizontal; D006801:Humans; D007231:Infant, Newborn; D020071:Interspersed Repetitive Sequences; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification; D008780:Methyltransferases / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D010957:Plasmids / Q000032:analysis* / Q000145:classification; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D047090:beta-Lactams / Q000494:pharmacology*
chemical_list: D000617:Aminoglycosides; D004269:DNA, Bacterial; D047090:beta-Lactams; D008780:Methyltransferases; C035181:rRNA (adenosine-O-2'-)methyltransferase