Detection of Staphylococcus aureus Delta-Toxin Production by Whole-Cell MALDI-TOF Mass Spectrometry

作者:Gagnaire Julie*; Dauwalder Olivier; Boisset Sandrine; Khau David; Freydiere Anne Marie; Ader Florence; Bes Michele; Lina Gerard; Tristan Anne; Reverdy Marie Elisabeth; Marchand Adrienne; Geissmann Thomas; Benito Yvonne; Durand Geraldine; Charrier Jean Philippe; Etienne Jerome; Welker Martin; Van Belkum Alex; Vandenesch Francois
来源:PLos One, 2012, 7(7): e40660.
DOI:10.1371/journal.pone.0040660

摘要

The aim of the present study was to detect the Staphylococcus aureus delta-toxin using Whole-Cell (WC) Matrix Assisted Laser Desorption Ionization - Time-of-Flight (MALDI-TOF) mass spectrometry (MS), correlate delta-toxin expression with accessory gene regulator (agr) status, and assess the prevalence of agr deficiency in clinical isolates with and without resistance to methicillin and glycopeptides. The position of the delta-toxin peak in the mass spectrum was identified using purified delta-toxin and isogenic wild type and mutant strains for agr-rnaIII, which encodes delta-toxin. Correlation between delta-toxin production and agr RNAIII expression was assessed by northern blotting. A series of 168 consecutive clinical isolates and 23 unrelated glycopeptide-intermediate S. aureus strains (GISA/heterogeneous GISA) were then tested by WC-MALDI-TOF MS. The delta-toxin peak was detected at 3005 +/- 5 Thomson, as expected for the naturally formylated delta toxin, or at 3035 +/- 5 Thomson for its G10S variant. Multivariate analysis showed that chronicity of S. aureus infection and glycopeptide resistance were significantly associated with delta-toxin deficiency (p = 0.048; CI 95%: 1.01-10.24; p = 0.023; CI 95%: 1.20-12.76, respectively). In conclusion, the S. aureus delta-toxin was identified in the WC-MALDI-TOF MS spectrum generated during routine identification procedures. Consequently, agr status can potentially predict infectious complications and rationalise application of novel virulence factor-based therapies.

  • 出版日期2012-7-6