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Mechanisms of Resistance

Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus

Duarte C. Oliveira, Hermínia de Lencastre
Duarte C. Oliveira
The Rockefeller University, New York, New York 10021, and Instituto de Tecnologia Química e Biológica, ITQB/UNL, 2780 Oeiras, Portugal
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Hermínia de Lencastre
The Rockefeller University, New York, New York 10021, and Instituto de Tecnologia Química e Biológica, ITQB/UNL, 2780 Oeiras, Portugal
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  • For correspondence: lencash@mail.rockefeller.edu
DOI: 10.1128/AAC.46.7.2155-2161.2002
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ABSTRACT

Full characterization of methicillin-resistant Staphylococcus aureus (MRSA) requires definition of not only the bacterial genetic background but also the structure of the complex and heterologous mec element these bacteria carry, which is associated with drug resistance determinant mecA. We report the development, validation, and application of a multiplex PCR strategy that allows quick presumptive characterization of the mec element types based on the structural features that were shown to be typical of mec elements carried by several MRSA clones. The strategy was validated by using a representative collection of pandemic MRSA clones in which the full structure of the associated mec elements was previously determined by hybridization and PCR screenings and also by DNA sequencing. The method was tested together with multilocus sequence typing and other typing methods for the characterization of 18 isolates representative of the MRSA clones recovered during a hospital outbreak in Barcelona, Spain. The multiplex PCR was shown to be rapid, robust, and capable in a single assay of identifying five structural types of the mec element among these strains, three major and two minor variants, each one of which has been already been seen among MRSA characterized earlier. This technique should be a useful addition to the armamentarium of molecular typing tools for the characterization of MRSA clonal types and for the rapid tentative identification of structural variants of the mec element.

  • Copyright © 2002 American Society for Microbiology
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Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus
Duarte C. Oliveira, Hermínia de Lencastre
Antimicrobial Agents and Chemotherapy Jul 2002, 46 (7) 2155-2161; DOI: 10.1128/AAC.46.7.2155-2161.2002

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Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus
Duarte C. Oliveira, Hermínia de Lencastre
Antimicrobial Agents and Chemotherapy Jul 2002, 46 (7) 2155-2161; DOI: 10.1128/AAC.46.7.2155-2161.2002
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KEYWORDS

Bacterial Proteins
Bacterial Typing Techniques
Carrier Proteins
Hexosyltransferases
methicillin resistance
Muramoylpentapeptide Carboxypeptidase
Peptidyl Transferases
Polymerase Chain Reaction
Staphylococcus aureus

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