title: Multiple drug resistant carbapenemases producing Acinetobacter baumannii isolates harbours multiple R-plasmids.
abstract: OBJECTIVE
  The nosocomial human pathogen Acinetobacter baumannii has high propensity to develop resistance to antimicrobials and to become multidrug resistant (MDR), consequently complicating the treatment. This study was carried out to investigate the presence of resistant plasmids (R-plasmids) among the clinical isolates of A. baumannii. In addition, the study was performed to check the presence of common β-lactamases encoding genes on these plasmids.
  
  
  METHODS
  A total of 55 clinical isolates of A. baumannii were included in the study and all were subjected to plasmid DNA isolation, followed by PCR to check the presence of resistance gene determinants such as blaOXA-23, blaOXA-51, blaOXA-58 and blaIMP-1 on these plasmids that encode for oxacillinase (OXA) and metallo-β-lactamase (MBL) type of carbapenemases. Plasmid curing experiments were carried out on selected isolates using ethidium bromide and acridine orange as curing agents and the antibiotic resistance profiles were evaluated before and after curing.
  
  
  RESULTS
  All the isolates were identified as A. baumannii by 16SrDNA amplification and sequencing. Plasmid DNA isolated from these isolates showed the occurrence of multiple plasmids with size ranging from 500bp to ≥25 kb. The percentage of blaOXA-51 and blaOXA-23 on plasmids were found to be 78 and 42 per cent, respectively and 20 isolates (36%) carried blaIMP-1 gene on plasmids. Significant difference was observed in the antibiograms of plasmid cured isolates when compared to their parental ones. The clinical isolates became susceptible to more than two antibiotic classes after curing of plasmids indicating plasmid borne resistance.
  
  
  CONCLUSIONS
  Our study determined the plasmid mediated resistance mechanisms and occurrence of different resistance genes on various plasmids isolated from MDR A. baumannii. The present findings showed the evidence for antibiotic resistance mediated through multiple plasmids in A. baumannii clinical isolates. This indicates towards a need for preventive measures to avert the dissemination of plasmid resistance determinants in clinical environments.
authors.forename: Rajagopalan
authors.initials: R
authors.lastname: Saranathan
authors.forename: Pagal
authors.initials: P
authors.lastname: Sudhakar
authors.affiliation: Department of Biotechnology, School of Life Sciences, Pondicherry University, Puducherry,India, .
authors.forename: R Uma
authors.initials: RU
authors.lastname: Karthika
authors.forename: Santosh Kumar
authors.initials: SK
authors.lastname: Singh
authors.forename: P
authors.initials: P
authors.lastname: Shashikala
authors.forename: Reba
authors.initials: R
authors.lastname: Kanungo
authors.forename: K
authors.initials: K
authors.lastname: Prashanth
mesh_terms: D040981:Acinetobacter baumannii / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D003428:Cross Infection / Q000382:microbiology*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics*; D006801:Humans; D007194:India; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011815:R Factors / Q000235:genetics*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D001618:beta-Lactamases; C063912:carbapenemase