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Related Experiment Videos

Microarray analysis of erythromycin resistance determinants.

D Volokhov1, V Chizhikov, K Chumakov

  • 1FDA Center for Food Safety and Applied Nutrition, College Park, MD 20740-3835, USA.

Journal of Applied Microbiology
|September 13, 2003
PubMed
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A new DNA microarray accurately detects macrolide, lincosamide, and streptogramin B (MLS) resistance genes in Staphylococcus aureus. This tool aids in identifying antibiotic resistance determinants across various bacterial strains.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Antibiotic resistance, particularly to macrolide, lincosamide, and streptogramin B (MLS) compounds, poses a significant threat in clinical settings.
  • Staphylococcus aureus is a major pathogen frequently associated with MLS resistance.

Purpose of the Study:

  • To develop a DNA microarray for the rapid and accurate analysis of genes conferring resistance to erythromycin and related MLS compounds.
  • To identify the prevalence of specific MLS resistance genes in clinical isolates of Staphylococcus aureus.

Main Methods:

  • Development of an oligonucleotide microarray with probes for six key MLS resistance genes (ermA, ermB, ermC, ereA, ereB, msrA/B).
  • Utilization of multiplex PCR for target gene amplification and fluorescent labeling from clinical bacterial strains.

Related Experiment Videos

  • Validation of microarray results through direct DNA sequencing.

    • Main Results:

    • The developed microarray successfully identified MLS resistance genes in reference and clinical strains of Staphylococcus aureus and Streptococcus pyogenes.
    • Eleven out of 18 tested S. aureus clinical isolates harbored MLS resistance determinants.
    • The ermC gene was present in all 11 resistant isolates, while ermA and msrA/B were found in over half of them. Multiple resistance genes were common, with 72% of resistant isolates carrying two or three genes.

    Conclusions:

    • Oligonucleotide microarrays provide a reliable method for detecting and identifying six major MLS resistance determinants.
    • Microarray-based detection of antibiotic resistance genes is a promising tool for identifying resistance mechanisms in diverse bacterial populations due to conserved gene homology.