Isolation and characterization of φkm18p, a novel lytic phage with therapeutic potential against extensively drug resistant Acinetobacter baumannii

PLoS One. 2012;7(10):e46537. doi: 10.1371/journal.pone.0046537. Epub 2012 Oct 5.

Abstract

Aims: To isolate phages against extensively drug resistant Acinetobacter baumannii (XDRAB) and characterize the highest lytic capability phage as a model to evaluate the potential on phage therapy.

Methods and results: Eight phages were isolated from hospital sewage and showed narrow host spectrum. Phage φkm18p was able to effectively lyse the most XDRAB. It has a dsDNA genome of 45 kb in size and hexagonal head of about 59 nm in diameter and no tail. Bacterial population decreased quickly from 10(8) CFU ml(-1) to 10(3) CFU ml(-1) in 30 min by φkm18p. The 185 kDa lysis protein encoded by φkm18p genome was detected when the extracted protein did not boil before SDS-PAGE; it showed that the lysis protein is a complex rather than a monomer. Phage φkm18p improved human lung epithelial cells survival rates when they were incubated with A. baumannii. Combination of phages (φkm18p, φTZ1 and φ314) as a cocktail could lyse all genotype-varying XDRAB isolates.

Conclusion: Infections with XDRAB are extremely difficult to treat and development of a phage cocktails therapy could be a therapeutic alternative in the future. Phage φkm18p is a good candidate for inclusion in phage cocktails.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acinetobacter Infections / microbiology
  • Acinetobacter Infections / therapy
  • Acinetobacter baumannii / drug effects
  • Acinetobacter baumannii / physiology
  • Acinetobacter baumannii / virology*
  • Anti-Bacterial Agents / pharmacology
  • Bacteriolysis*
  • Bacteriophages / genetics
  • Bacteriophages / isolation & purification
  • Bacteriophages / physiology*
  • Biological Therapy
  • Cell Line
  • Cell Survival
  • DNA, Viral / genetics
  • Drug Resistance, Bacterial*
  • Endopeptidases / metabolism
  • Genome, Viral
  • Humans
  • Restriction Mapping
  • Sewage / virology
  • Viral Proteins / metabolism
  • Viral Tropism

Substances

  • Anti-Bacterial Agents
  • DNA, Viral
  • Sewage
  • Viral Proteins
  • Endopeptidases
  • endolysin

Grants and funding

This work was supported by grants from the National Science Council, Republic of China (No. NSC 94-2311-B-214-001) and I-Shou University (ISU93-02-17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.