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Related Experiment Videos

Reproducibility of Staphylococcus epidermidis plasmid profiles.

A I Hartstein1, V H Morthland, A L Rashad

  • 1Department of Hospital Infection Control, University Hospitals, Portland, Oregon.

Diagnostic Microbiology and Infectious Disease
|July 1, 1991
PubMed
Summary

Staphylococcus epidermidis plasmid profiles are reproducible when stored below -20°C. Room temperature storage leads to plasmid DNA loss, impacting profile stability and analysis.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacteriology

Background:

  • Plasmid profiling is crucial for bacterial identification and tracking.
  • Staphylococcus epidermidis is an opportunistic pathogen with significant clinical relevance.
  • Understanding factors affecting plasmid stability is essential for reliable microbiological studies.

Purpose of the Study:

  • To assess the reproducibility of Staphylococcus epidermidis plasmid profiles over time.
  • To determine the impact of storage temperature and extraction methods on plasmid stability.
  • To identify optimal conditions for maintaining plasmid profiles for accurate analysis.

Main Methods:

  • Subculturing and storage of six Staphylococcus epidermidis isolates at -70°C, -20°C, and room temperature for 8 months.

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  • Plasmid DNA extraction using two methods: mixed alkyltrimethylammonium bromide (ATAB) and modified Parisi (Brij 58 and deoxycholate).
  • Analysis of plasmid DNA via electrophoresis and restriction endonuclease digestion.

    • Main Results:

    • Plasmid DNA loss occurred in isolates stored at room temperature.
    • Plasmid profiles were generally reproducible across different storage conditions and extraction methods, except for room temperature storage.
    • Minor variations in high molecular weight plasmid DNA bands were observed but were not consistently reproducible.
    • The modified Parisi extraction method showed unreliable results with restriction endonuclease digestion.

    Conclusions:

    • Staphylococcus epidermidis plasmid profiles are highly reproducible when isolates are stored at or below -20°C.
    • Storage at room temperature compromises plasmid stability and profile accuracy.
    • Minor discrepancies in large plasmid DNA bands can occur but are manageable with repetitive testing or restriction analysis of ATAB-extracted DNA.