Use of chemiluminescent DNA probes in the rapid detection of oxacillin resistance in clinically isolated strains of Staphylococcus aureus

Gary R. Youmans, Thomas Davis, DeAnna D. Fuller

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Chemiluminescent DNA probes (AccuProbe, species specific; and FlashTrack, bacterial generic) were used to determine oxacillin resistance in Staphylococcus aureus. Ribosomal RNA was measured at designated intervals in the presence and absence of antibiotic. A total of 48 (AccuProbe assay) and 24 (FlashTrack) S. aureus isolates with known oxacillin susceptibility patterns were inoculated into Bactec 6A bottles both with and without 4 μg/ml oxacillin and incubated at 35°C for 4 h. Aliquots were removed at 0 and 4 h, and pellets of bacteria were obtained via selective centrifugation. Probe assay counts (relative light units, RLUs) were performed. Of 21 oxacillin-resistant S. aureus (ORSA) strains, 20 showed a >5-fold RLU increase during the incubation period (AccuProbe assay): 25 of 27 oxacillin-susceptible strains demonstrated a ≤4-fold increase. AccuProbe test sensitivity and specificity were 95% and 92%, respectively. With the generic FlashTrack probe assay, all nine ORSA isolates showed a ≥4-to 10-fold increase in RLUs, and all 15 oxacillin-susceptible strains showed a ≤4-fold increase in RLUs during the 4-h incubation. The FlashTrack test sensitivity and specificity were both 100%. Probe assays were completed within 5 h. This study suggests that rapid and reliable determination of oxacillin resistance in S. aureus clinical isolates can be accomplished using commercially available DNA probes.

Original languageEnglish
Pages (from-to)99-104
Number of pages6
JournalDiagnostic Microbiology and Infectious Disease
Volume16
Issue number2
DOIs
StatePublished - 1993
Externally publishedYes

Fingerprint

Oxacillin
DNA Probes
Staphylococcus aureus
Light
Sensitivity and Specificity
Ribosomal RNA
Centrifugation
Anti-Bacterial Agents
Bacteria

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology
  • Microbiology
  • Parasitology
  • Virology
  • Immunology and Allergy
  • Infectious Diseases

Cite this

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title = "Use of chemiluminescent DNA probes in the rapid detection of oxacillin resistance in clinically isolated strains of Staphylococcus aureus",
abstract = "Chemiluminescent DNA probes (AccuProbe, species specific; and FlashTrack, bacterial generic) were used to determine oxacillin resistance in Staphylococcus aureus. Ribosomal RNA was measured at designated intervals in the presence and absence of antibiotic. A total of 48 (AccuProbe assay) and 24 (FlashTrack) S. aureus isolates with known oxacillin susceptibility patterns were inoculated into Bactec 6A bottles both with and without 4 μg/ml oxacillin and incubated at 35°C for 4 h. Aliquots were removed at 0 and 4 h, and pellets of bacteria were obtained via selective centrifugation. Probe assay counts (relative light units, RLUs) were performed. Of 21 oxacillin-resistant S. aureus (ORSA) strains, 20 showed a >5-fold RLU increase during the incubation period (AccuProbe assay): 25 of 27 oxacillin-susceptible strains demonstrated a ≤4-fold increase. AccuProbe test sensitivity and specificity were 95{\%} and 92{\%}, respectively. With the generic FlashTrack probe assay, all nine ORSA isolates showed a ≥4-to 10-fold increase in RLUs, and all 15 oxacillin-susceptible strains showed a ≤4-fold increase in RLUs during the 4-h incubation. The FlashTrack test sensitivity and specificity were both 100{\%}. Probe assays were completed within 5 h. This study suggests that rapid and reliable determination of oxacillin resistance in S. aureus clinical isolates can be accomplished using commercially available DNA probes.",
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AU - Fuller, DeAnna D.

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