Solid-phase microbead array for multiplex O-serotyping of Escherichia coli
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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.
KeywordsBacteria Screening Fluorescence Diagnosis Automation Imaging Image processing High-throughput Coating
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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