Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF/MS)-based identification of pathogens from positive blood culture bottles

Abstract

Since the expansion of commercial use of MALDI-TOF/MS instruments for the identification of bacteria from culture which has occurred over the past 5-8 years, techniques for the identification of bacteria directly from positive blood cultures have been developed (Lagace-Wiens et al., J Clin Microbiol 50:3324-3328, 2012; Martiny et al., Eur J Clin Microbiol Infect Dis 31:2269-2281, 2012; Moussaoui et al., Clin Microbiol Infect 16:1631-1638, 2010). These techniques have the potential to provide definitive identification of pathogens causing sepsis 18-48 h earlier than conventional methodologies, and implementation of these methods has been shown to impact morbidity and hospital costs in a positive way (Martiny et al., Clin Microbiol Infect 19:E568-E581, 2013; Loonen et al., Eur J Clin Microbiol Infect Dis 31:1575-1583, 2012). Although many methods for purification of bacterial cells have been developed, including differential centrifugation, centrifuge lysis, and preincubation on sold media (March-Rossello et al., Eur J Clin Microbiol Infect Dis 32:699-704, 2013; Saffert et al., Diagn Microbiol Infect Dis 73:21-26, 2012; Schubert et al., J Mol Diagn 13:701-706, 2011), we will describe the method by which intact bacterial cells are extracted from positive blood culture bottles using a commercially available kit (SepsiTyper™) which is based on the centrifuge lysis methodology (Lagace-Wiens et al., J Clin Microbiol 50:3324-3328, 2012; Buchan et al., J Clin Microbiol 50:346-352, 2012).