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PCR-based ribotyping of Staphylococcus aureus.

A M Oliveira1, M C Ramos

  • 1Disciplina de Moléstias Infecciosas e Parasitárias, Departamento de Clínica Médica, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Campinas, SP, Brasil. alemacedo@uol.com.br

Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Medicas E Biologicas
|February 16, 2002
PubMed
Summary

PCR-based ribotyping offers a simpler alternative to pulsed-field gel electrophoresis for typing Staphylococcus aureus. This method effectively distinguishes methicillin-sensitive strains, revealing diverse hospital-wide distribution, but is less effective for methicillin-resistant strains.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Epidemiology

Background:

  • Staphylococcus aureus infections necessitate effective genotyping for hospital epidemiology.
  • Pulsed-field gel electrophoresis (PFGE) is the standard but is labor-intensive and costly.
  • Alternative, more accessible genotyping methods are needed.

Purpose of the Study:

  • To evaluate PCR-based ribotyping using the 16S-23S rRNA gene region as a genotyping tool for Staphylococcus aureus.
  • To compare the discriminatory power of PCR-based ribotyping with PFGE.

Main Methods:

  • Genomic DNA was analyzed using PCR with three primer pairs targeting the 16S-23S rRNA region.
  • Agar gel electrophoresis and ethidium bromide staining were used to visualize amplified DNA fragments.

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  • The method's resolution was tested on clinical isolates, including samples from a single patient, known diverse strains, and a large collection of hospital-acquired strains.

    • Main Results:

    • PCR-based ribotyping correctly identified identical strains from the same patient.
    • The technique differentiated between strains with known distinct PFGE patterns.
    • It revealed a polyclonal distribution of methicillin-sensitive S. aureus (22 unique patterns from 27 strains).
    • However, it showed limited diversity among methicillin-resistant S. aureus isolates (one pattern from 61 strains).

    Conclusions:

    • PCR-based ribotyping is a valuable, accessible technique for genotyping methicillin-sensitive Staphylococcus aureus.
    • The method is less effective for characterizing methicillin-resistant strains due to limited polymorphism.
    • This technique can aid in understanding the epidemiology of S. aureus in hospital settings.