S. Wesley Long, Randall J. Olsen, Shrenik C. Mehta, Timothy Palzkill, Patricia L. Cernoch, Katherine K. Perez, William L. Musick, Adriana E. Rosato, James M. Musser.
Antimicrobial Agents and Chemotherapy. 2014. Vol 58.

Overview

  • Keywords: Genome sequencing, antibiotic resistance, clinical isolates, ceftaroline
  • Aim of the study: Understanding antibiotic resistance mechanism in clinical isolates of Staphylococcus aureus
  • Application: Genome sequencing
  • Sample type: Patient expectorated sputum & blood
  • Material: FastPrep-96™ Instrument, Lysing Matrix B tubes
  • Buffer: Tryptic soy broth

Protocols & Parameters

  1. Patient isolates were grown on tryptic soy agar supplemented with 5% sheep blood.
  2. Five of the isolates grew from expectorated sputum. And the sixth isolate was obtained from an aerobic blood culture bottle.
  3. Genomic DNA was isolated from multiple colonies grown overnight in tryptic soy broth.
  4. The cells were lysed using Lysing Matrix B in a in a FastPrep-96™ instrument.

Conclusion

  • The use of the high-throughput FastPrep-96™ homogenizer in combination with Lysing Matrix B tubes allows high quality DNA extraction and genome sequencing analysis of ceftaroline-resistant methicillin-resistant Staphylococcus aureus (MRSA).
  • Genome sequencing results confirm a previously undescribed high-level antibiotic resistance mechanism in clinical isolates of MRSA.