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Raman spectroscopic detection of physiology changes in plasmid-bearing Escherichia coli with and without antibiotic treatment

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Abstract

Bacterial resistances against antibiotics are increasingly problematic for medical treatment of pathogenic bacteria, e.g., in hospitals. Resistances are, among other genes, often encoded on plasmids which can be transmitted between bacteria not only within one species, but also between different species, genera, and families. The plasmid pDrive is transformed into bacteria of the model strain Escherichia coli DH5α. Within this investigation, we applied micro-Raman spectroscopy with two different excitation wavelengths in combination with support vector machine (SVM) and linear discriminant analysis (LDA) to differentiate between bacterial cultures according to their cultural plasmid content. Recognition rates of about 92% and 90% are achieved by Raman excitation at 532 and 244 nm, respectively. The SVM loadings reveal that the pDrive transformed bacterial cultures exhibit a higher DNA content compared to the untransformed cultures. To elucidate the influence of the antibiotic, ampicillin-treated cultures are also comprised within this study and are classified with rates of about 97% and 100% for 532 and 244 nm Raman excitation, respectively. The Raman spectra recorded with 532 nm excitation wavelength show differences of the secondary protein structure and enhanced stress-related respiration rates for the ampicillin-treated cultures. Independent cultural replicates of either ampicillin-challenged or non-challenged cultures are successfully identified with identification rates of over 90%.

The plasmid content of bacteria is directly evaluated by means of Raman spectroscopy

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Acknowledgment

We gratefully acknowledge financial support from the Deutsche Forschungsgemeinschaft (Graduiertenkolleg GK 1257 “Alteration and element mobility at the microbe-mineral interface” in the frame of the Jena School of Microbial Communication and Po563/7-2).

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Authors and Affiliations

  1. Institut für Physikalische Chemie, Friedrich-Schiller-Universität, Helmholtzweg 4, 07743, Jena, Germany

    Angela Walter, Thomas Bocklitz, Wilm Schumacher, Petra Rösch & Jürgen Popp

  2. Institut für Mikrobiologie, Friedrich-Schiller-Universität, Neugasse 25, 07743, Jena, Germany

    Martin Reinicke & Erika Kothe

  3. Institut für Photonische Technologien, Albert-Einstein-Strasse 9, 07745, Jena, Germany

    Jürgen Popp

Authors
  1. Angela Walter
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  2. Martin Reinicke
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  3. Thomas Bocklitz
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  4. Wilm Schumacher
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  5. Petra Rösch
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  6. Erika Kothe
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  7. Jürgen Popp
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Correspondence to Jürgen Popp.

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Published in the special issue Biophotonics with Guest Editors Jürgen Popp and Reiner Salzer.

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Walter, A., Reinicke, M., Bocklitz, T. et al. Raman spectroscopic detection of physiology changes in plasmid-bearing Escherichia coli with and without antibiotic treatment. Anal Bioanal Chem 400, 2763–2773 (2011). https://doi.org/10.1007/s00216-011-4819-4

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  • DOI: https://doi.org/10.1007/s00216-011-4819-4

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