Discrimination of bacterial strains by Fourier-transform near-infrared spectroscopy using an aluminum oxide membrane

Luis E. Rodriguez-Saona; Farukh M. Khambaty; Francis S. Fry; Elizabeth M. Calvey

doi:10.1117/12.455148

7 February 2002 Discrimination of bacterial strains by Fourier-transform near-infrared spectroscopy using an aluminum oxide membrane

Luis E. Rodriguez-Saona, Farukh M. Khambaty, Francis S. Fry, Elizabeth M. Calvey

Author Affiliations +

Proceedings Volume 4574, Instrumentation for Air Pollution and Global Atmospheric Monitoring; (2002) https://doi.org/10.1117/12.455148
Event: Environmental and Industrial Sensing, 2001, Boston, MA, United States

Abstract

To address the need for a fast and sensitive method for the detection of bacterial contamination in solutions, the use of Fourier-transform near infrared (FT-NIR) spectroscopy and multivariate pattern recognition techniques was evaluated. The complex cellular composition of bacteria yields FT-NIR vibrational transitions (overtone and combination bands) that might be useful for identification and sub-typing. Bacteria including strains of Escherichia coli spp., Pseudomonas aeruginosa, Bacillus spp. and Listeria innocua were evaluated. The harvested cells were treated with ethanol (70% v/v) to reduce the safety concerns when evaluating pathogenic strains. The bacterial cells were concentrated on an aluminum oxide membrane to obtain a thin bacterial film. Spectra were collected by FT-NIR by using a diffuse reflection-integrating sphere. This simple membrane filtration procedure generated reproducible FT-NIR spectra that can be used for rapid discrimination among closely related strains. Principal Component Analysis (PCA) of transformed spectra in the 5000-4000 cm^-1 region exhibited clusters that discriminated between bacteria species at levels < 1 mg wet cells weight (approximately 10⁶-10⁷ CFU/mg). Variations in the growth conditions of the bacteria substantially affected the FT-NIR spectra and diminished the ability of PCA to differentiate among strains; this underscores the importance of developing robust sampling protocols. FT-NIR in conjunction with multivariate techniques can be used for the rapid and accurate evaluation of potential bacterial contamination in liquids with minimal sample manipulation.

Citation Download Citation

Luis E. Rodriguez-Saona, Farukh M. Khambaty, Francis S. Fry, and Elizabeth M. Calvey "Discrimination of bacterial strains by Fourier-transform near-infrared spectroscopy using an aluminum oxide membrane", Proc. SPIE 4574, Instrumentation for Air Pollution and Global Atmospheric Monitoring, (7 February 2002); https://doi.org/10.1117/12.455148

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