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Replication of the Ordered, Nonredundant Library of Pseudomonas aeruginosa strain PA14 Transposon Insertion Mutants
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Genomic analysis reveals multi-lineage carbapenem-resistant Pseudomonas aeruginosa mimicking a hospital outbreak

Süleyman Yalçın1, Sezin Ünlü Çelebi2, Salih Rıza Ozan Yıldız3

  • 1Ministry of Health, General Directorate of Public Health, Microbiology Reference Laboratory, Ankara, Türkiye.

BMC Microbiology
|July 17, 2026
PubMed
Abstract

Insights

Multidrug-resistant Pseudomonas aeruginosa outbreaks in hospitals can involve multiple distinct bacterial clones, not just a single strain. Whole-genome sequencing is crucial for understanding transmission and guiding infection control for these complex nosocomial infections.

Area of Science:

  • Clinical microbiology and infectious diseases.
  • Genomics and molecular epidemiology.
  • Hospital infection control and prevention.

Background:

  • Pseudomonas aeruginosa is a significant cause of hospital-acquired infections, frequently displaying multidrug resistance (MDR) and adaptability.
  • This study examined a cluster of MDR P. aeruginosa isolates from five hospitalized patients, noting similar resistance patterns suggestive of a common source.
  • Pulsed-field gel electrophoresis (PFGE) revealed four distinct clones, indicating clonal heterogeneity rather than a single-strain outbreak.

Purpose of the Study:

  • To investigate the genetic diversity and molecular basis of antimicrobial resistance and virulence in a cluster of MDR P. aeruginosa isolates from a single healthcare facility.
  • To determine if the observed temporal cluster represented a single-strain outbreak or the co-circulation of multiple P. aeruginosa clones.
  • To leverage whole-genome sequencing (WGS) for a comprehensive understanding of resistance mechanisms, virulence factors, and genomic relationships.

Main Methods:

  • Isolation and characterization of MDR P. aeruginosa from five hospitalized patients.
  • Antimicrobial susceptibility testing to determine resistance profiles.
  • Pulsed-field gel electrophoresis (PFGE) for initial strain typing.
  • Whole-genome sequencing (WGS) for in-depth genomic analysis, including resistome and virulome profiling.

Main Results:

  • All isolates exhibited resistance to multiple antibiotics (e.g., carbapenems, fluoroquinolones) but remained susceptible to colistin.
  • Resistome analysis identified diverse antimicrobial resistance genes, including beta-lactamases (blaPDC, blaOXA, blaVIM-2) and various efflux pumps.
  • Virulome analysis detected determinants for biofilm formation, adherence, motility, immune evasion, and toxin production, with evidence of horizontal gene transfer (HGT).

Conclusions:

  • Nosocomial outbreaks of MDR P. aeruginosa can involve multiple unrelated clones co-circulating, challenging traditional outbreak investigation assumptions.
  • Despite similar resistance profiles, the presence of four distinct clones highlights that such events are not always caused by a single strain.
  • WGS is essential for accurately dissecting transmission dynamics in MDR P. aeruginosa outbreaks and informing effective infection control strategies.
Keywords:
Pseudomonas aeruginosaClonal HeterogeneityGenotypingNosocomial outbreakWhole-genome sequencing

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