Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus. (c2013)

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dc.contributor.author El Hage, Hussein Hassan
dc.date.accessioned 2015-02-10T09:51:27Z
dc.date.available 2015-02-10T09:51:27Z
dc.date.issued 2015-02-10
dc.date.submitted 2013-06-11
dc.identifier.uri http://hdl.handle.net/10725/1917
dc.description Includes bibliographical references (leaves 39-44). en_US
dc.description.abstract Matrix Assisted Laser Desorption Ionization mass spectrometry (MALDI-MS) has been recently introduced in to the field of microbiology. The use of this technique overcomes some of the limitations of current phenotypic and genotypic methods. MALDI-typing of microorganisms is an approach based on the differentiation of MALDI-acquired protein fingerprints. Employed in clinical settings, it allows rapid identification of microorganisms down to the strain level. Leading to high morbidity and mortality rates, antibiotic resistant strains of Staphylococcus aureus have become a worldwide concern. Herein, we carried out a comparative study of 20 variations of an acid/alcohol bacterial protein extraction method using a clinical isolate of S. aureus. Protein fingerprints of these extracts acquired in linear mode (800-20,000 Da) were used for assessment of information content (number of peaks) and identity (size of proteins/peptides, m/z). Two methods were shown to be most efficient for sample preparation, i.e. formic acid/methanol and trifluoroacetic acid/ethanol, yielding 28 peaks each. Proteins obtained by the classic formic acid/ethanol extraction were separated using 2-dimensional electrophoresis (2-DE) yielding more than 50 protein spots. Nine proteins were successfully identified using peptide mass fingerprinting (PMF), two of which are virulence related proteins, SpoVG and endonuclease IV. These proteins are considered prospects for MALDI-typing. en_US
dc.language.iso en en_US
dc.subject Staphylococcus aureus -- Identification en_US
dc.subject Matrix-assisted laser desorption-ionization en_US
dc.subject Proteins -- Analysis en_US
dc.subject Lebanese American University -- Dissertations en_US
dc.subject Dissertations, Academic en_US
dc.title Optimization of sample preparation method, and evaluation of formic acid protein extracts for MALDI-typing of Staphylococcus aureus. (c2013) en_US
dc.type Thesis en_US
dc.term.submitted Spring en_US
dc.author.degree MS in Molecular Biology en_US
dc.author.school Arts and Sciences en_US
dc.author.idnumber 200902489 en_US
dc.author.commembers Dr. Roy khalaf
dc.author.commembers Dr. Georges Khazen
dc.author.woa OA en_US
dc.description.physdesc 1 hard copy: xv, 44, [25] leaves; 30 cm. available at RNL. en_US
dc.author.division Biology en_US
dc.author.advisor Dr. Brigitte Wex
dc.keywords Staphylococcus aureus en_US
dc.keywords Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS) en_US
dc.keywords Protein Fingerprint en_US
dc.keywords MALDI-Typing en_US
dc.keywords 2-Dimensional Electrophoresis (2-DE) en_US
dc.keywords Peptide Mass Fingerprinting (PMF) en_US
dc.identifier.doi https://doi.org/10.26756/th.2013.38 en_US
dc.publisher.institution Lebanese American University en_US

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