This work presents a polymeric centrifugal microfluidic platform for the rapid and sensitive identification of bacteria directly from urine, thus eliminating time-consuming cultivation steps. This “Lab-on-a-Disc” platform utilizes the rotationally induced centrifugal field to efficiently capture bacteria directly from suspension within a glass-polymer hybrid chip. Once trapped in an array of small V-shaped structures, the bacteria are readily available for spectroscopic characterization, such as Raman spectroscopic fingerprinting, providing valuable information on the characteristics of the captured bacteria. Utilising fluorescence microscopy, quantification of the bacterial load has been achieved for concentrations above 2 × 10−7 cells ml−1 within a 4 μl sample. As a pilot application, we characterize urine samples from patients with urinary tract infections. Following minimal sample preparation, Raman spectra of the bacteria are recorded following centrifugal capture in stopped-flow sedimentation mode. Utilizing advanced analysis algorithms, including extended multiplicative scattering correction, high-quality Raman spectra of different pathogens, such as Escherichia coli or Enterococcus faecalis, are obtained from the analyzed patient samples. The whole procedure, including sample preparation, requires about 1 h to obtain a valuable result, marking a significant reduction in diagnosis time when compared to the 24 h and more typically required for standard microbiological methods. As this cost-efficient centrifugal cartridge can be operated using low-complexity, widely automated instrumentation, while providing valuable bacterial identification in urine samples in a greatly reduced time-period, our opto-microfluidic Lab-on-a-Disc device demonstrates great potential for next-generation patient diagnostics at the of point-of-care.
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Rapid, culture-independent, optical diagnostics of centrifugally captured bacteria from urine samples
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July 2015
Research Article|
August 11 2015
Rapid, culture-independent, optical diagnostics of centrifugally captured bacteria from urine samples
Ulrich-Christian Schröder
;
Ulrich-Christian Schröder
a)
1Integrated Research and Treatment Center, “Center for Sepsis Control and Care” (CSCC),
Jena University Hospital
, Erlanger Allee 101, 07747 Jena, Germany
2
Leibniz Institute of Photonic Technology Jena e.V.
, 07749 Jena, Germany
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Frank Bokeloh;
Frank Bokeloh
a)
1Integrated Research and Treatment Center, “Center for Sepsis Control and Care” (CSCC),
Jena University Hospital
, Erlanger Allee 101, 07747 Jena, Germany
3Biomedical Diagnostics Institute, National Centre of Sensor Research, School of Physical Sciences,
Dublin City University
, Glasnevin, Dublin 9, Ireland
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Mary O'Sullivan;
Mary O'Sullivan
3Biomedical Diagnostics Institute, National Centre of Sensor Research, School of Physical Sciences,
Dublin City University
, Glasnevin, Dublin 9, Ireland
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Uwe Glaser;
Uwe Glaser
1Integrated Research and Treatment Center, “Center for Sepsis Control and Care” (CSCC),
Jena University Hospital
, Erlanger Allee 101, 07747 Jena, Germany
2
Leibniz Institute of Photonic Technology Jena e.V.
, 07749 Jena, Germany
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Katharina Wolf;
Katharina Wolf
4Institute of Medical Microbiology,
Jena University Hospital
, 07747 Jena, Germany
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Wolfgang Pfister;
Wolfgang Pfister
4Institute of Medical Microbiology,
Jena University Hospital
, 07747 Jena, Germany
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Jürgen Popp;
Jürgen Popp
1Integrated Research and Treatment Center, “Center for Sepsis Control and Care” (CSCC),
Jena University Hospital
, Erlanger Allee 101, 07747 Jena, Germany
2
Leibniz Institute of Photonic Technology Jena e.V.
, 07749 Jena, Germany
5Institute of Physical Chemistry and Abbe Center of Photonics,
Friedrich Schiller University Jena
, 07743 Jena, Germany
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Jens Ducrée
;
Jens Ducrée
a)
3Biomedical Diagnostics Institute, National Centre of Sensor Research, School of Physical Sciences,
Dublin City University
, Glasnevin, Dublin 9, Ireland
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Ute Neugebauer
Ute Neugebauer
1Integrated Research and Treatment Center, “Center for Sepsis Control and Care” (CSCC),
Jena University Hospital
, Erlanger Allee 101, 07747 Jena, Germany
2
Leibniz Institute of Photonic Technology Jena e.V.
, 07749 Jena, Germany
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a)
U.-C. Schröder, F. Bokeloh, J. Ducrée, and U. Neugebauer contributed equally to this work.
b)
Author to whom correspondence should be addressed. Electronic mail: ute.neugebauer@med.uni-jena.de. Tel.: +49-3641-9323364, Fax: +49-3641-9323382.
Biomicrofluidics 9, 044118 (2015)
Article history
Received:
May 08 2015
Accepted:
July 24 2015
Citation
Ulrich-Christian Schröder, Frank Bokeloh, Mary O'Sullivan, Uwe Glaser, Katharina Wolf, Wolfgang Pfister, Jürgen Popp, Jens Ducrée, Ute Neugebauer; Rapid, culture-independent, optical diagnostics of centrifugally captured bacteria from urine samples. Biomicrofluidics 1 July 2015; 9 (4): 044118. https://doi.org/10.1063/1.4928070
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