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Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
09:03

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus

Published on: September 5, 2013

Rapid microarray-based identification of different mecA alleles in Staphylococci.

Stefan Monecke1, Elke Müller, Stefan Schwarz

  • 1Institute for Medical Microbiology and Hygiene, Technical University of Dresden, Dresden, Germany. monecke@rocketmail.com

Antimicrobial Agents and Chemotherapy
|August 15, 2012
PubMed
Summary

The study identified diverse mecA alleles in Staphylococcus species, with only some conferring methicillin resistance. This highlights the need to consider mecA allele variability in diagnostic test design.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • The mecA gene is crucial for methicillin resistance in Staphylococcus species.
  • Existing diagnostic methods may not fully capture the diversity of mecA alleles.
  • Understanding mecA allele variation is essential for accurate detection of antibiotic resistance.

Purpose of the Study:

  • To analyze the diversity of mecA alleles.
  • To develop a microarray for rapid discrimination of mecA alleles.
  • To investigate the association between mecA alleles and beta-lactam resistance.

Main Methods:

  • Bioinformatic analysis of published mecA sequences.
  • Design and application of a custom microarray assay.
  • Characterization of clinical and veterinary Staphylococcus isolates.
  • Determination of Minimum Inhibitory Concentrations (MICs) for beta-lactam antibiotics.

Main Results:

  • Analysis revealed 32 distinct mecA alleles clustering into 15 hybridization patterns on the microarray.
  • The assay identified 9 expected and 1 novel pattern in 78 Staphylococcus isolates.
  • mecA alleles associated with beta-lactam resistance were linked to only 4 of the 9 observed patterns.
  • mecA alleles not conferring resistance were mainly found in coagulase-negative staphylococci of animal origin.

Conclusions:

  • Significant diversity exists among mecA alleles, impacting their association with beta-lactam resistance.
  • A substantial number of mecA alleles do not confer resistance, particularly in certain animal-associated staphylococci.
  • Diagnostic assays for mecA detection must account for sequence variability and differential effects on antibiotic susceptibility.