Differentiation of bacterial spores via 2D-IR spectroscopy

Procacci, Barbara and Rutherford, Samantha H. and Greetham, Gregory M. and Towrie, Michael and Parker, Anthony W. and Robinson, Camilla V. and Howle, Christopher R. and Hunt, Neil T. (2021) Differentiation of bacterial spores via 2D-IR spectroscopy. Spectrochimica Acta - Part A: Molecular and Biomolecular Spectroscopy, 249. 119319. ISSN 1386-1425 (https://doi.org/10.1016/j.saa.2020.119319)

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Abstract

Ultrafast 2D-IR spectroscopy is a powerful tool for understanding the spectroscopy and dynamics of biological molecules in the solution phase. A number of recent studies have begun to explore the utility of the information-rich 2D-IR spectra for analytical applications. Here, we report the application of ultrafast 2D-IR spectroscopy for the detection and classification of bacterial spores. 2D-IR spectra of Bacillus atrophaeus and Bacillus thuringiensis spores as dry films on CaF2 windows were obtained. The sporulated nature of the bacteria was confirmed using 2D-IR diagonal and off-diagonal peaks arising from the calcium dipicolinate CaDP·3H2O biomarker for sporulation. Distinctive peaks, in the protein amide I region of the spectrum were used to differentiate the two types of spore. The identified marker modes demonstrate the potential for the use of 2D-IR methods as a direct means of spore classification. We discuss these new results in perspective with the current state of analytical 2D-IR measurements, showing that the potential exists to apply 2D-IR spectroscopy to detect the spores on surfaces and in suspensions as well as in dry films. The results demonstrate how applying 2D-IR screening methodologies to spores would enable the creation of a library of spectra for classification purposes.

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