Abstract

CHARACTERIZATION OF MICROORGANISMS OF INTEREST TO HOMELAND SECURITY AND PUBLIC HEALTH UTILIZING LIQUID CHROMATOGRAPHY/MASS SPECTROMETRY By: Robert Anthony Everley, Ph.D. A Dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Chemistry at Virginia Commonwealth University. Virginia Commonwealth University, 2008 Major Director: Timothy R. Croley Adjunct Professor, Department of Chemistry 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. Automated charge state deconvolution, spectral subtraction and spectral mirroring were used to reveal subtle differences in the LC/MS data. Reproducible intact protein biomarkers were discovered based on their unique mass, retention time and relative intensity. These markers were implemented to differentiate closely related strain types, (e.g. two distinct isolates of E. coli O157:H7) and to correctly identify unknown pathogens. Notable, was the distinction of multiple serotypes of enterohaemorrhagic E. coli which cannot be distinguished by clinical

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