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12 February 2014 Direct identification of clinically relevant bacterial and yeast microcolonies and macrocolonies on solid culture media by Raman spectroscopy
Isabelle Espagnon, Denis Ostrovskii, Raphael Mathey, Mathieu G. Dupoy, Pierre Joly, Armelle Novelli-Rousseau, Fredric Pinston, Olivier Gal, Frédéric Mallard, Denis F. Leroux
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Journal of Biomedical Optics, Vol. 19, Issue 2, 027004 (February 2014). https://doi.org/10.1117/1.JBO.19.2.027004
Abstract
Decreasing turnaround time is a paramount objective in clinical diagnosis. We evaluated the discrimination power of Raman spectroscopy when analyzing colonies from 80 strains belonging to nine bacterial and one yeast species directly on solid culture medium after 24-h (macrocolonies) and 6-h (microcolonies) incubation. This approach, that minimizes sample preparation and culture time, would allow resuming culture after identification to perform downstream antibiotic susceptibility testing. Correct identification rates measured for macrocolonies and microcolonies reached 94.1% and 91.5%, respectively, in a leave-one-strain-out cross-validation mode without any correction for possible medium interference. Large spectral differences were observed between macrocolonies and microcolonies, that were attributed to true biological differences. Our results, conducted on a very diversified panel of species and strains, were obtained by using simple and robust sample preparation and preprocessing procedures, while still confirming published results obtained by using more complex elaborated protocols. Instrumentation is simplified by the use of 532-nm laser excitation yielding a Raman signal in the visible range. It is, to our knowledge, the first side-by-side full classification study of microorganisms in the exponential and stationary phases confirming the excellent performance of Raman spectroscopy for early species-level identification of microorganisms directly from an agar culture.
CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
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Isabelle Espagnon, Denis Ostrovskii, Raphael Mathey, Mathieu G. Dupoy, Pierre Joly, Armelle Novelli-Rousseau, Fredric Pinston, Olivier Gal, Frédéric Mallard, and Denis F. Leroux "Direct identification of clinically relevant bacterial and yeast microcolonies and macrocolonies on solid culture media by Raman spectroscopy," Journal of Biomedical Optics 19(2), 027004 (12 February 2014). https://doi.org/10.1117/1.JBO.19.2.027004
Published: 12 February 2014
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Cited by 37 scholarly publications and 6 patents.
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KEYWORDS
Raman spectroscopy
Yeast
Signal to noise ratio
Databases
Solids
Bacteria
Principal component analysis
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