Instrumental improvements and sample preparations that enable reproducible, reliable acquisition of mass spectra from whole bacterial cells. (27th September 2015)

• Record Type: Journal Article
• Authors: Alusta, Pierre; Buzatu, Dan; Williams, Anna; Cooper, Willie‐Mae; Tarasenko, Olga; Dorey, R. Cameron; Hall, Reggie; Parker, W. Ryan; Wilkes, Jon G.
• Is Part Of: Rapid communications in mass spectrometry. Volume 29:Number 21(2015)
• Page Start: 1961
• Page End: 1968
• Publication Date: 2015-09-27
• Subjects: Mass spectrometry -- Periodicals; 543.65
• DOI: 10.1002/rcm.7299
• Languages: English
• ISSNs: 0951-4198

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="rcm7299-sec-0001" sec-type="section"> <title>Rationale</title> <p>Rapid sub‐species characterization of pathogens is required for timely responses in outbreak situations. Pyrolysis mass spectrometry (PyMS) has the potential to be used for this purpose.</p> </sec> <sec id="rcm7299-sec-0002" sec-type="section"> <title>Methods</title> <p>However, in order to make PyMS practical for traceback applications, certain improvements related to spectrum reproducibility and data acquisition speed were required. The main objectives of this study were to facilitate fast detection (<30 min to analyze 6 samples, including preparation) and sub‐species‐level bacterial characterization based on pattern recognition of mass spectral fingerprints acquired from whole cells volatilized and ionized at atmospheric pressure. An AccuTOF DART mass spectrometer was re‐engineered to permit ionization of low‐volatility bacteria by means of <italic>Plasma Jet Ionization</italic> (PJI), in which an electric discharge, and, by extension, a plasma beam, impinges on sample cells.</p> </sec> <sec id="rcm7299-sec-0003" sec-type="section"> <title>Results</title> <p>Instrumental improvements and spectral acquisition methodology are described. Performance of the re‐engineered system was assessed using a small challenge set comprised of assorted bacterial isolates differing in identity by varying amounts. In general, the spectral patterns obtained allowed differentiation of all samples tested, including those of the same genus and species but different serotypes.</p> </sec> <sec id="rcm7299-sec-0004" sec-type="section"> <title>Conclusions</title> <p>Fluctuations of ±15% in bacterial cell concentrations did not substantially compromise replicate spectra reproducibility. © 2015 National Center for Toxicological Research. <italic>Rapid Communications in Mass Spectrometry</italic> published by John Wiley & Sons Ltd.</p> </sec> </abstract>