Rapid lysis of gram-positive cocci for pulsed-field gel electrophoresis using achromopeptidase

Rebecca B. Leonard, Karen C. Carroll

Research output: Contribution to journalArticlepeer-review

Abstract

To adapt pulsed-field gel electrophoresis (PFGE) to a busy clinical laboratory, we evaluated the use of achromopeptidase (ACP) as the sole agent for lysis of a variety of gram-positive cocci. Growth in an appropriate broth media was adjusted to a 3.0 McFarland density before aliquotting for washing and plug preparation. ACP incorporated into the agarose plugs, or added to the buffer solution, lysed cells in <1 h when suspended in a Tris-NaCl buffer. Two 30-rain washes in a Tris-EDTA buffer were sufficient to remove cellular debris without additional deproteination. Final PFGE results were obtained in -48 h, including the initial broth subculturing.

Original languageEnglish (US)
Pages (from-to)288-291
Number of pages4
JournalDiagnostic Molecular Pathology
Volume6
Issue number5
DOIs
StatePublished - Oct 1 1997
Externally publishedYes

Keywords

  • Achromopeptidase
  • Molecular epidemiology
  • Pulsed-field gel electrophoresis

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

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