Evaluation of fluorescence-based amplified fragment length polymorphism analysis for molecular typing in hospital epidemiology

Comparison with pulsed-field gel electrophoresis for typing strains of vancomycin-resistant Enterococcus faecium

N. A. Antonishyn, R. R. McDonald, E. L. Chan, G. Horsman, C. E. Woodmansee, P. S. Falk, C. G. Mayhall

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Fluorescence-based amplified fragment length polymorphism (fbAFLP) is a novel assay based on the fluorescent analysis of an amplified subset of restriction fragments. The fbAFLP assay involves the selective PCR amplification of restriction fragments from a total digest of genomic DNA. The ligation of adapters with primer-specific sites coupled with primers containing selective nucleotides allowed the full potential of PCR to be realized while maintaining the advantages of restriction endonuclease analysis. Fluorescence-based fragment analysis with polyacrylamide gel electrophoresis provides the accurate band sizing required for homology assessment. The large number of phylogenetically informative characters obtained by fbAFLP is well suited for cluster analysis and database development. The method demonstrated excellent reproducibility and ease of performance and interpretation. We typed 30 epidemiologically well-characterized isolates of vancomycin-resistant enterococci from an outbreak in a university hospital by fbAFLP. Clustering of fbAFLP data matched epidemiological, microbiological, and pulsed-field gel electrophoresis data. This study demonstrates the unprecedented utility of fbAFLP for epidemiological investigation. Future developments in standardization and automation will set fbAFLP as the "gold standard" for molecular typing in epidemiology.

Original languageEnglish (US)
Pages (from-to)4058-4065
Number of pages8
JournalJournal of Clinical Microbiology
Volume38
Issue number11
StatePublished - 2000
Externally publishedYes

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Amplified Fragment Length Polymorphism Analysis
Molecular Typing
Enterococcus faecium
Pulsed Field Gel Electrophoresis
Epidemiology
Fluorescence
Cluster Analysis
Polymerase Chain Reaction
Vancomycin-Resistant Enterococci
Automation
DNA Restriction Enzymes
Disease Outbreaks
Ligation
Polyacrylamide Gel Electrophoresis
Nucleotides
Databases

ASJC Scopus subject areas

  • Microbiology (medical)
  • Microbiology

Cite this

Evaluation of fluorescence-based amplified fragment length polymorphism analysis for molecular typing in hospital epidemiology : Comparison with pulsed-field gel electrophoresis for typing strains of vancomycin-resistant Enterococcus faecium. / Antonishyn, N. A.; McDonald, R. R.; Chan, E. L.; Horsman, G.; Woodmansee, C. E.; Falk, P. S.; Mayhall, C. G.

In: Journal of Clinical Microbiology, Vol. 38, No. 11, 2000, p. 4058-4065.

Research output: Contribution to journalArticle

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