Microchimica Acta

, Volume 184, Issue 5, pp 1405–1415 | Cite as

Solid-phase microbead array for multiplex O-serotyping of Escherichia coli

  • Claudia Liebsch
  • Stefan Rödiger
  • Alexander Böhm
  • Jörg Nitschke
  • Jörg Weinreich
  • Angelika Fruth
  • Dirk Roggenbuck
  • Werner Lehmann
  • Uwe Schedler
  • Thomas Juretzek
  • Peter Schierack
Original Paper

Abstract

Typing and classification of Escherichia coli (E. coli) according to cell wall components, like polysaccharides, is routinely done by serotyping. Given the presence of 188 known O-antigens, this process is complex. The authors present a proof-of-concept planar microbead array for multiplexed O-serotyping. Ten clinically relevant E. coli serotypes associated with high risk for diarrhea in humans were examined (O26, O55, O78, O118, O124, O127, O128, O142, O145 and O157). Antisera were assigned to specific microbead populations, which can be differentiated by size and fluorescence color. Automatted image processing and data analysis were conducted by a microscopic interpretation platform. Homogenous antiserum coating of the microbeads was demonstrated by an intra-population CV that ranges from 3.3 to 6.3% and by an inter-population CV of 9.5%. Typical detections limits are in the range from 0.31 to 0.71 refMFI. Significantly elevated fluorescence signals revealed that E. coli of a certain serogroup bound specifically to microbeads with the matching antiserum (p < 0.001). In our perception, the method represents a viable diagnostic tool for automated multiplex serotyping of E. coli. It enables simultaneous and high-throughput screening for different O-antigens by a simple staining and binding protocol.

Graphical abstract

Schematic of a planar microbead array for the typing and classification of E. coli according to cell wall components. Based on coated fluorescent microbeads, multiplex O-serotyping of E. coli is accomplished via fluorescence imaging.

Keywords

Bacteria Screening Fluorescence Diagnosis Automation Imaging Image processing High-throughput Coating 

Notes

Acknowledgements

We thank Michael Schenker and Jörg Weinreich for excellent technical assistance. This work was supported by InnoProfile 03IP611, by InnoProfile-Transfer 03IPT611A and 03IPT611X funded by the Federal Ministry of Education and Research (BMBF, Germany).

Compliance with ethical standards

Dirk Roggenbuck has a management role and is a shareholder of GA Generic Assays GmbH. This company is a diagnostic manufacturer. Werner Lehmann has a management role and is a shareholder of Attomol GmbH. This company is a diagnostic manufacturer. Uwe Schedler has a management role and is a shareholder of PolyAn GmbH. This company is a supplier for diagnostic manufacturers. All other authors declare that they have no competing financial interests.

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Copyright information

© Springer-Verlag Wien 2017

Authors and Affiliations

  • Claudia Liebsch
    • 1
  • Stefan Rödiger
    • 1
  • Alexander Böhm
    • 1
  • Jörg Nitschke
    • 1
  • Jörg Weinreich
    • 1
  • Angelika Fruth
    • 2
  • Dirk Roggenbuck
    • 1
    • 3
  • Werner Lehmann
    • 4
  • Uwe Schedler
    • 5
  • Thomas Juretzek
    • 6
  • Peter Schierack
    • 1
  1. 1.Institute of BiotechnologyBrandenburg University of Technology Cottbus-SenftenbergSenftenbergGermany
  2. 2.Branch WernigerodeRobert Koch-InstituteWernigerodeGermany
  3. 3.GA Generic Assays GmbHDahlewitzGermany
  4. 4.Attomol GmbHLiptenGermany
  5. 5.PolyAn GmbHBerlinGermany
  6. 6.Carl-Thiem-Klinikum CottbusCottbusGermany

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