title: Applying a PCR-based open-reading frame typing method for easy genotyping and molecular epidemiological analysis of Pseudomonas aeruginosa.
abstract: OBJECTIVE
  Molecular epidemiological techniques, such as pulsed-field gel electrophoresis (PFGE), or multilocus sequence typing (MLST) have facilitated our understanding of the transmission routes of nosocomial infections by Pseudomonas aeruginosa. However, they are time consuming and technically demanding. To perform molecular epidemiological analysis in a standard microbiology laboratory, we aimed to develop a simpler and effective molecular epidemiological technique based on the open-reading frame (ORF) distribution patterns detected by PCR, which we call PCR-based ORF typing (POT).
  
  
  RESULTS
  Ten ORFs from genomic islets, five ORFs from genomic islands, and the metallo-β-lactamases (MBLs) blaIMP and blaVIM were selected by comparing the whole-genome sequences of different Ps. aeruginosa strains (PAO1, PA7, UCBPP-PA14 and LESB58). These 17 ORFs were detected, along with a Ps. aeruginosa marker, using 9-plex and 10-plex PCR systems. The genotypes in the POT were compared to those obtained by using PFGE and MLST.
  
  
  CONCLUSIONS
  Using the POT method, molecular epidemiological analyses of Ps. aeruginosa can be completed in 4 h.
  
  
  CONCLUSIONS
  Since this method is very easy to perform, even in standard clinical laboratories, it could be a valuable tool for monitoring daily infection control measures.
authors.affiliation: Laboratory of Bacteriology, Aichi Prefectural Institute of Public Health, Nagoya, Japan.|Department of Infectious Diseases and Immunology, Clinical Research Center, National Hospital Organization Nagoya Medical Center, Nagoya, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Laboratory of Bacteriology, Aichi Prefectural Institute of Public Health, Nagoya, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.affiliation: Laboratory of Bacteriology, Aichi Prefectural Institute of Public Health, Nagoya, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Aoki
authors.affiliation: Department of Infectious Diseases and Immunology, Clinical Research Center, National Hospital Organization Nagoya Medical Center, Nagoya, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Hosoba
authors.affiliation: Laboratory of Bacteriology, Aichi Prefectural Institute of Public Health, Nagoya, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Matsumoto
authors.affiliation: Department of Infectious Diseases, Kanazawa Medical University, Ishikawa, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Baba
authors.affiliation: Department of Infectious Diseases, Kanazawa Medical University, Ishikawa, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Iinuma
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D003428:Cross Infection / Q000453:epidemiology / Q000382:microbiology*; D044404:Genomic Islands; D005838:Genotype; D006801:Humans; D017720:Molecular Epidemiology / Q000379:methods*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction / Q000379:methods*; D011552:Pseudomonas Infections / Q000453:epidemiology / Q000382:microbiology*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification*; D001618:beta-Lactamases / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D001618:beta-Lactamases
keywords: PCR; Pseudomonas aeruginosa; genotyping; molecular epidemiology; rapid method