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


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Article Info

DATE

2017-05

AUTHORS

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

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 abstractSchematic 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. 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. More... »

PAGES

1405-1415

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00604-017-2088-4

DOI

http://dx.doi.org/10.1007/s00604-017-2088-4

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1084022884


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