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Related Experiment Videos

A novel gentamicin resistance gene in Enterococcus

J W Chow1, M J Zervos, S A Lerner

  • 1Department of Internal Medicine, Wayne State University School of Medicine, Detroit, Michigan, USA.

Antimicrobial Agents and Chemotherapy
|March 1, 1997
PubMed
Summary

A novel gentamicin resistance gene, aph(2")-Ic, was identified in Enterococcus gallinarum SF9117. This gene may lead to underdetection of high-level gentamicin resistance in clinical settings.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Antimicrobial Resistance

Background:

  • Enterococcus gallinarum SF9117 exhibits high-level gentamicin resistance (MIC of 256 µg/mL).
  • Standard combination therapy with ampicillin and gentamicin did not demonstrate synergistic effects against this isolate.
  • Initial molecular investigation using a probe for aac(6 olyl)-aph(2") did not yield positive hybridization with SF9117 DNA.

Purpose of the Study:

  • To identify the genetic basis of gentamicin resistance in Enterococcus gallinarum SF9117.
  • To characterize the novel gentamicin resistance gene and its prevalence in other enterococcal species.

Main Methods:

  • Cloning of a 3.2-kb fragment from plasmid pYN134 of SF9117 into Escherichia coli DH5α.
  • Nucleotide sequence analysis of the cloned fragment to identify open reading frames.

Related Experiment Videos

  • Homology searches and functional characterization of the deduced protein sequence.
  • Hybridization studies using an intragenic probe with clinical isolates of Enterococcus faecium and Enterococcus faecalis.

    • Main Results:

    • A 918-bp open reading frame, designated aph(2")-Ic, was identified, encoding a protein with homology to bifunctional AAC(6 olyl)-APH(2") enzymes.
    • The cloned fragment conferred gentamicin resistance to E. coli DH5α, and the enzyme activity was provisionally named APH(2")-Ic.
    • The aph(2")-Ic gene was detected in genomic DNA of an Enterococcus faecium isolate and plasmid DNA of an Enterococcus faecalis isolate.

    Conclusions:

    • A novel gentamicin resistance gene, aph(2")-Ic, has been identified in Enterococcus gallinarum.
    • This gene confers high-level gentamicin resistance and may be present in other clinically relevant enterococcal species.
    • Current clinical laboratory screening methods for high-level gentamicin resistance may fail to detect isolates harboring aph(2")-Ic, potentially impacting patient treatment.