Automated laser fluorescence analysis of randomly amplified polymorphic DNA: a rapid method for investigating nosocomial transmission of Acinetobacter baumannii

J Med Microbiol. 1995 Dec;43(6):446-51. doi: 10.1099/00222615-43-6-446.

Abstract

A rapid method for genotyping Acinetobacter baumannii based on PCR-fingerprinting with fluorescent primers was evaluated. Automated laser fluorescence analysis (ALFA) enabled on-line generation of high resolution DNA-fingerprints during polyacrylamide gel electrophoresis of randomly amplified polymorphic DNA (RAPD) products. The results were in concordance with macro-restriction fragment patterns produced by pulse-field gel electrophoresis (PFGE) of ApaI digests of chromosomal DNA. RAPD-ALFA was able to identify homologous strains suggestive of horizontal transmission in < 8 h after colonies were obtained on solid media, whereas PFGE analysis took c. 90 h. Speed and digitised data format renders RAPD-ALFA attractive for routine in-house epidemiological screening of isolates from intensive care and other hospital units.

MeSH terms

  • Acinetobacter / classification
  • Acinetobacter / genetics*
  • Acinetobacter Infections / epidemiology
  • Acinetobacter Infections / microbiology
  • Acinetobacter Infections / transmission*
  • Base Sequence
  • Chromosomes, Bacterial / genetics
  • Cross Infection / epidemiology
  • Cross Infection / microbiology
  • Cross Infection / transmission*
  • DNA Fingerprinting / methods
  • DNA Primers / chemistry
  • DNA, Bacterial / analysis*
  • Disease Outbreaks
  • Electrophoresis, Gel, Pulsed-Field
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescence
  • Fluorescent Dyes
  • Gene Amplification / genetics*
  • Genotype
  • Germany / epidemiology
  • Humans
  • Lasers
  • Molecular Sequence Data

Substances

  • DNA Primers
  • DNA, Bacterial
  • Fluorescent Dyes