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Updated: Jul 11, 2026

Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
Published on: September 5, 2013
[Comparison of phenotypic methods with PCR to screening methicillin resistance in coagulase negative staphylococci]
D Majouri1, A Touati, W Achour
1Laboratoire de microbiologie, centre national de greffe de moelle osseuse, rue Djebel-Lakhdhar, Bab Saadoun, 1006 Tunis, Tunisie.
The Aim Of Study:
Appreciation of the frequency, the level and the genetic support of methicillin resistance.
Material And Methods:
Seventy-three strains of coagulase negative staphylococci isolated from various specimens, from January to June 2004, were studied. The phenotypic detection was carried out by disk diffusion test using oxacillin and cefoxitin disks, by the determination of oxacillin Minimal Inhibitor Concentration (E-test), by the oxacillin screening test at a concentration of 4 mug/ml and by the search of the penicillin binding protein PBP2a using the slide latex agglutination test. The results of these methods were compared to PCR of mecA gene.
Results:
Forty-eight strains carried mecA gene whose 30 were detected by the oxacillin disk, the cefoxitin disk, the oxacillin screening test, the slide latex agglutination test and had a MIC from 24 to 256 mug/ml. Seventeen strains were not detected by oxacillin disk but by cefoxitin disk and the slide latex agglutination test. Among these strains, 13 (76%) had oxacillin MIC from 0.5 to 1,5 mug/ml and not grew on oxacillin agar screening, while 4 (24%) had oxacillin MIC from 6 to 16 mug/ml and grew on this agar. One strain had oxacillin MIC of 0,19 mug/ml and was not detected with any phenotypic method.
Conclusion:
The determination of oxacillin MIC, the search of the PBP2a or more simply the cefoxitin disk had permitted to detect the strains mecA gene (+) with resistant and pre-resistant phenotype but not the strain with sensible phenotype (2.1%).
Insights
Methicillin resistance in staphylococci was assessed using various methods. Cefoxitin disk and PBP2a detection effectively identified mecA gene-positive strains, highlighting their utility in clinical settings.
Area of Science:
- Microbiology
- Genetics
- Clinical Diagnostics
Context:
- Coagulase-negative staphylococci (CoNS) are significant opportunistic pathogens.
- Methicillin resistance in staphylococci poses a major challenge in healthcare settings.
- Accurate detection of methicillin resistance is crucial for effective treatment and infection control.
Purpose:
- To evaluate the frequency, level, and genetic basis of methicillin resistance in CoNS.
- To compare the efficacy of various phenotypic detection methods against PCR detection of the mecA gene.
- To assess the performance of oxacillin and cefoxitin disks, oxacillin Minimal Inhibitor Concentration (MIC), oxacillin screening tests, and PBP2a detection.
Summary:
- Seventy-three CoNS strains were analyzed. PCR confirmed the presence of the mecA gene in 48 strains.
- Phenotypic methods, including cefoxitin disk and PBP2a detection, showed high concordance with mecA gene presence.
- Some strains with low-level or heterogeneous resistance were challenging to detect with certain phenotypic tests.
Impact:
- Cefoxitin disk testing and PBP2a detection are reliable methods for identifying methicillin-resistant staphylococci, including those with pre-resistant phenotypes.
- Understanding detection limitations is vital for accurate diagnosis and patient management.
- This study contributes to optimizing antimicrobial susceptibility testing protocols for staphylococcal infections.
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