Liquid chromatography/mass spectrometry characterization of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Shigella</Emphasis> species |
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Authors: | Robert A Everley Tiffany M Mott Shane A Wyatt Denise M Toney Timothy R Croley |
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Institution: | (1) Biology Department, San Francisco State University, 1600 Holloway Ave, San Francisco, CA 94132, USA;(2) Pathology, Microbiology & Immunology Department, School of Veterinary Medicine, University of California, Davis, CA 95616, USA;(3) Monterey County Public Health Department, 1270 Natividad Rd, Salinas, CA 93906, USA |
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Abstract: | Liquid chromatography/quadrupole time of flight mass spectrometry (LC/QTOF MS) utilizing electrospray ionization was employed
to monitor protein expression in Escherichia coli and Shigella organisms. Comparison with MALDI/TOF-MS revealed more proteins, particularly above 15 kDa. A combination of automated charge
state deconvolution, spectral mirroring, and spectral subtraction was used to reveal subtle differences in the LC/MS data.
Reproducible intact protein biomarker candidates were discovered based on their unique mass, retention time, and relative
intensity. These marker candidates were implemented to differentiate closely related strain types, (e.g., two distinct isolates
of E. coli O157:H7) and to correctly identify unknown pathogens. This LC/MS approach is less labor-intensive than pulsed-field gel electrophoresis,
affords greater specificity than real-time PCR, and requires no primers or antibodies. Additionally, this approach would be
beneficial during outbreaks of foodborne disease or bioterrorism investigations by complementing methods typically used in
diagnostic microbiology laboratories. |
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