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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

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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.