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Development of a modified DNA extraction method for pulsed-field gel electrophoresis analysis of Staphylococcus

Mohammad Mehdi Feizabadi1, Arash Ghodousi, Bizhan Nomanpour

  • 1Department for Microbiology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. mfeizabadi@tums.ac.ir

Journal of Microbiological Methods
|November 17, 2010
PubMed

Insights

A new pulsed-field gel electrophoresis (PFGE) method for Staphylococcus aureus and enterococci eliminates lysostaphin, significantly reducing costs for bacterial typing. This optimized protocol makes PFGE more accessible for clinical diagnostics.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Clinical Diagnostics

Background:

  • Pulsed-field gel electrophoresis (PFGE) is a crucial technique for bacterial strain typing.
  • Lysostaphin is commonly used in PFGE protocols for Staphylococcus aureus and enterococci but adds significant cost.
  • Reducing reagent expenses is vital for widespread clinical application of PFGE.

Purpose of the Study:

  • To develop a modified PFGE protocol for Staphylococcus aureus and enterococci.
  • To eliminate the need for costly lysostaphin in the spheroplast formation step.
  • To reduce the overall cost of PFGE typing for these clinically relevant bacteria.

Main Methods:

  • A modified pulsed-field gel electrophoresis (PFGE) protocol was developed.
  • The protocol was applied to clinical isolates of Staphylococcus aureus and enterococci.
  • Lysostaphin was omitted during the spheroplast formation stage.

Main Results:

  • The modified PFGE protocol successfully typed clinical isolates of Staphylococcus aureus and enterococci.
  • The elimination of lysostaphin significantly reduced the cost associated with PFGE typing.
  • The protocol maintained the effectiveness of spheroplast formation without lysostaphin.

Conclusions:

  • A cost-effective modified PFGE protocol for Staphylococcus aureus and enterococci has been established.
  • This method reduces expenses by removing the requirement for lysostaphin.
  • The optimized protocol enhances the accessibility of PFGE for routine clinical use.

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