title: Chemometric studies for the characterization and differentiation of microorganisms using in situ derivatization and thermal desorption ion mobility spectrometry.
abstract: Whole-cell bacteria were characterized and differentiated by thermal desorption ion mobility spectrometry and chemometric modeling. Principal component analysis was used to evaluate the differences in the ion mobility spectra of whole-cell bacteria and the fatty acid methyl esters (FAMEs) generated in situ after derivatization of the bacterial lipids. Alternating least squares served to extract bacterial peaks from the complex ion mobility spectra of intact microorganisms and, therefore, facilitated the characterization of bacterial strains, species, and Gram type. In situ thermal hydrolysis/methylation with tetramethylammonium hydroxide was necessary for the differentiation of Escherichia coli strains, which otherwise could not be distinguished by spectra acquired with the ITEMISER ion mobility spectrometer. The addition of the methylating agent had no effect on Gram-positive bacteria, and therefore, they could not be differentiated by genera. The classification of E. coli strains was possible by analysis of the IMS spectra from the FAMEs generated in situ. By using the fuzzy multivariate rule-building expert system and cross-validation, a correct classification rate of 96% (22 out of 23 spectra) was obtained. Chemometric modeling on bacterial ion mobility spectra coupled to thermal hydrolysis/methylation proved a simple, rapid (2 min/sample), inexpensive, and sensitive technique to characterize and differentiate intact microorganisms. The ITEMISER ion mobility spectrometer could detect as few as 4 x 10(6) cells/sample.
authors.affiliation: Center for Intelligent Chemical Instrumentation, Department of Chemistry and Biochemistry, Clippinger Laboratories, Ohio University, Athens, OH 45701-2979, USA.
authors.forename: Mariela L
authors.initials: ML
authors.lastname: Ochoa
authors.forename: Peter B
authors.initials: PB
authors.lastname: Harrington
mesh_terms: D004926:Escherichia coli / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000145:classification / Q000302:isolation & purification / Q000378:metabolism; D006868:Hydrolysis; D008745:Methylation; D008828:Microbiological Techniques; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D013057:Spectrum Analysis / Q000379:methods*; D013696:Temperature; D013997:Time Factors
chemical_list: D000644:Quaternary Ammonium Compounds; C027917:tetramethylammonium