Express fluorescence in situ hybridization methods for the detection of Staphylococcus aureus

Clin Lab. 2011;57(9-10):789-94.

Abstract

Background: As a proof-of-concept, the feasibility of detecting Staphylococcus aureus faster than previous whole-cell fluorescent in situ hybridization (FISH) methods was tested.

Methods: Isolates of Staphylococcus were treated with three rapid slide-based FISH protocols and DNA probes. Protocols were shortened by optimizing, combining or omitting steps.

Results: All FISH protocols detected S. aureus and not the phenotypically similar Staphylococcus epidermidis. The express FISH assay was completed in 24 minutes. The one-step FISH assay with NaCl and the one-step with phosphate buffered saline (PBS) assay took 19 minutes, but yielded a weaker signal.

Conclusions: The exploratory study identified S. aureus two to three times faster than previous methods. Additional testing in a clinical laboratory scenario (for example with positive blood-culture bottles) is warranted.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA, Bacterial / analysis
  • High-Throughput Screening Assays
  • Humans
  • In Situ Hybridization, Fluorescence / methods*
  • Staphylococcal Infections / diagnosis*
  • Staphylococcal Infections / microbiology
  • Staphylococcus aureus / cytology
  • Staphylococcus aureus / genetics
  • Staphylococcus aureus / isolation & purification*
  • Staphylococcus epidermidis / genetics
  • Staphylococcus epidermidis / isolation & purification
  • Time Factors

Substances

  • DNA, Bacterial