Staphylococcus aureus Specific FRET Probe-Based Antibacterial Susceptibility Testing (SF-AST) by Detection of Micrococcal Nuclease Activity

ACS Infect Dis. 2020 Feb 14;6(2):215-223. doi: 10.1021/acsinfecdis.9b00260. Epub 2020 Jan 29.

Abstract

In this study, we describe a simple and rapid antibacterial susceptibility testing (AST) method for Staphylococcus aureus called S. aureus specific fluorescence resonance energy transfer (FRET) probe-based AST (SF-AST), which is based on an S. aureus specific FRET probe (SF probe) that detects micrococcal nuclease (MNase) activity secreted from S. aureus. The SF-AST was tested with an S. aureus quality control (QC) strain against six relevant antibiotics, and the minimum inhibitory concentration (MIC) values obtained with the broth microdilution (BMD) method were compared, as a gold standard AST. Results were obtained with high accuracy in 4-6 h. The MIC for the methicillin resistance using 20 clinical S. aureus isolates of SF-AST showed 100% sensitivity, specificity, positive predictive value, and negative predictive value, as compared to BMD. Thus, the SF-AST method is a simple, rapid, and useful antibiotic resistance test for S. aureus, and it provides a basis for clinical treatment in a short time.

Keywords: Staphylococcus aureus; antibacterial susceptibility testing; fluorescence resonance energy transfer; micrococcal nuclease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • DNA Probes
  • Fluorescence Resonance Energy Transfer
  • Methicillin Resistance
  • Microbial Sensitivity Tests
  • Micrococcal Nuclease / metabolism*
  • Predictive Value of Tests
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Staphylococcus aureus / drug effects*
  • Staphylococcus aureus / enzymology*

Substances

  • Anti-Bacterial Agents
  • DNA Probes
  • Micrococcal Nuclease