Affiliation:
1. Institute of Physical Chemistry, Friedrich Schiller University Jena, Helmholtzweg 4
2. Friedrich Loeffler Institute, Federal Research Institute for Animal Health, Institute of Bacterial Infections and Zoonoses, Naumburger Straße 96a, D-07743 Jena
3. Institute of Photonic Technology, Albert-Einstein-Straße 9, D-07745 Jena, Germany
Abstract
ABSTRACT
Micro-Raman spectroscopy is a fast and sensitive tool for the detection, classification, and identification of biological organisms. The vibrational spectrum inherently serves as a fingerprint of the biochemical composition of each bacterium and thus makes identification at the species level, or even the subspecies level, possible. Therefore, microorganisms in areas susceptible to bacterial contamination, e.g., clinical environments or food-processing technology, can be sensed. Within the scope of point-of-care-testing also, detection of intentionally released biosafety level 3 (BSL-3) agents, such as
Bacillus anthracis
endospores, or their products is attainable. However, no Raman spectroscopy-compatible inactivation method for the notoriously resistant
Bacillus
endospores has been elaborated so far. In this work we present an inactivation protocol for endospores that permits, on the one hand, sufficient microbial inactivation and, on the other hand, the recording of Raman spectroscopic signatures of single endospores, making species-specific identification by means of highly sophisticated chemometrical methods possible. Several physical and chemical inactivation methods were assessed, and eventually treatment with 20% formaldehyde proved to be superior to the other methods in terms of sporicidal capacity and information conservation in the Raman spectra. The latter fact has been verified by successfully using self-learning machines (such as support vector machines or artificial neural networks) to identify inactivated
B. anthracis
-related endospores with adequate accuracies within the range of the limited model database employed.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
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