Authors: Samuels, Alan C.¹; Snyder, A. Peter¹; Emge, Darren K.¹; Amant, Diane S.T.²; Minter, Jennifer³; Campbell, Mark⁴; Tripathi, Ashish⁵

Source: Applied Spectroscopy, Volume 63, Issue 1, Pages 6A-30A and 1-131 (January 2009) , pp. 14-24(11)

Publisher: Society for Applied Spectroscopy

Abstract:

Fourier transform infrared (FT-IR) spectroscopy historically is a powerful tool for the taxonomic classification of bacteria by genus, species, and strain when they are grown under carefully controlled conditions. Relatively few reports have investigated the determination and classification of pathogens such as the National Institute of Allergy and Infectious Diseases (NIAID) Category A Bacillus anthracis spores and cells (BA), Yersinia species, Francisella tularensis (FT), and Category B Brucella species from FT-IR spectra. We investigated the multivariate statistics classification ability of the FT-IR spectra of viable pathogenic and non-pathogenic NIAID Category A and B bacteria. The impact of different growth media, growth time and temperature, rolling circle filter of the data, and wavelength range were investigated for their microorganism differentiation capability. Viability of the bacteria was confirmed by agar plate growth after the FT-IR experimental procedures were performed. Principal component analysis (PCA) was reduced to maps of two PC vectors in order to distill the FT-IR spectral features into manageable, visual presentations. The PCA results of the strains of BA, FT, Brucella, and Yersinia spectra from conditions of varying growth media and culture time were readily separable in two-dimensional (2D) PC plots. FT spectra were separated from those of the three other genera. The BA pathogenic spore strains 1029, LA1, and Ames were clearly differentiated from the rest of the dataset. Yersinia rhodei, Y. enterocolitica, and Y. pestis species were distinctly separated from the remaining dataset and could also be classified by growth media. Different growth media produced distinct subsets in the FT, BA, and Yersinia spp. regions in the 2D PC plots. Various 2D PC plots provided differential degrees of separation with respect to the four viable bacterial genera including the BA sub-categories of pathogenic spores, vegetative cells, and nonpathogenic vegetative cells. This work provided evidence that FT-IR spectroscopy can indeed separate the four major pathogenic bacterial genera of NIAID Category A and B biological threat agents including details according to the growth conditions and statistical parameters.

Keywords: FOURIER TRANSFORM INFRARED SPECTROSCOPY; FT-IR SPECTROSCOPY; BACTERIA DISCRIMINATION; YERSINIA; BACILLUS ANTHRACIS; BRUCELLA; FRANCISELLA TULARENSIS; PRINCIPAL COMPONENT ANALYSIS; PCA; BIOLOGICAL DISCRIMINATION; GROWTH MEDIA; GROWTH TIMES; WAVELENGTH RANGES; ROLLING CIRCLE FILTER; SPORES

Document Type: Research article

DOI: http://dx.doi.org/10.1366/000370209787169867

Affiliations: 1: ECBC, Research and Technology Directorate, Edgewood Area, Aberdeen Proving Ground, Maryland 21010-5424 2: Camber Corporation, Edgewood Area, SFAE-CBD-GN-C, Aberdeen Proving Ground, Maryland 21010-5424 3: Science and Technology Corporation, Edgewood, Maryland 21040 4: Science and Technology Corporation, Edgewood, Maryland 21040; US Department of Agriculture, Athens, GA 30605 5: Science Applications International Corporation, Research and Technology Directorate, Aberdeen Proving Ground, Maryland 21010-5424

Publication date: 2009-01-01

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