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Related Experiment Videos

PCR fragment length polymorphism analysis of vancomycin-resistant Enterococcus faecium.

S Donabedian1, E Hershberger, L A Thal

  • 1Departments of Medicine, Wayne State University, Detroit, Michigan, USA.

Journal of Clinical Microbiology
|August 2, 2000
PubMed
Summary

This study found that the Tn1546 element, responsible for glycopeptide resistance in VanA Enterococcus faecium, is frequently altered in clinical isolates. Most isolates showed changes in the vanSH region, indicating widespread dissemination of modified resistance genes.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • VanA Enterococcus faecium is a significant cause of hospital-acquired infections.
  • Glycopeptide resistance in Enterococcus faecium is often mediated by the Tn1546 element.
  • Understanding variations in Tn1546 is crucial for infection control.

Purpose of the Study:

  • To investigate the genetic variations of the Tn1546 glycopeptide resistance element in clinical isolates of VanA Enterococcus faecium.
  • To determine the prevalence of different Tn1546 variants among isolates from Michigan hospitals.

Main Methods:

  • Utilized PCR fragment length polymorphism to analyze the vanA gene cluster within the Tn1546 element.
  • Employed pulsed-field gel electrophoresis (PFGE) to assess the genetic relatedness of isolates.

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  • Compared PCR amplicons to a reference Tn1546 element from E. faecium 228.

    • Main Results:

    • Five distinct PCR fragment length patterns were identified, indicating alterations in Tn1546.
    • A significant majority (79.2%) of isolates exhibited an insertion between the vanS and vanH genes, resulting in a larger vanSH amplicon.
    • Other variations included insertions within the vanS gene and the vanYZ region.

    Conclusions:

    • The Tn1546 element is frequently altered in VanA Enterococcus faecium clinical isolates.
    • The prevalence of an insertion between vanS and vanH suggests the dissemination of a common, modified Tn1546 element.
    • These findings highlight the dynamic nature of antimicrobial resistance genes in hospital settings.