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

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

doi:10.1007/s00604-017-2088-4

Cited by 8 publications

(7 citation statements)

References 35 publications

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“…The labeled biomolecules as well as the labeled microbeads were obtained from commercial sources. The VideoScan technology is a fluorescence microscopy-based technology developed in our laboratory and is described elsewhere. ,, …”

Section: Methodsmentioning

confidence: 99%

“…The VideoScan technology is a fluorescence microscopy-based technology developed in our laboratory and is described elsewhere. 7,21,22 Microbead Functionalization and Detection. Carboxylated poly(methyl methacrylate) microbeads (PolyAn, Berlin) were used.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…Assays based on color coding introduce different dyes to either the targets or the substrate and hence discriminate several probes in a single testing. For instance, fluorescence microbeads of different sizes and equipped with dyes are distinguished by flow cytometry, microfluidics, or with fluorescence microscopy-based detection platforms. − …”

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confidence: 99%

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Spatial Separation of Microbeads into Detection Levels by a Bioorthogonal Porous Hydrogel for Size-Selective Analysis and Increased Multiplexity

et al. 2019

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Multiplex detection techniques are emerging within the fields of life science research and medical diagnostics where it is mandatory to analyze a great number of molecules. The detection techniques need to be highly efficient but often involve complicated and expensive fabrication procedures. Here, we present the immobilization and geometric separation of fluorescence-labeled microbeads for a multiplex detection in k levels. A compound of differently sized target molecules (DNA, proteins) is channeled into the respective detection levels by making use of a hydrogel as a size selective filter. The immobilized microbeads (10–20 μm) are considerably larger than the pores of the hydrogel network and therefore stay fixed at the well bottom and in higher elevations, respectively. Small biomolecules can diffuse through the pores of the network, whereas medium-sized biomolecules pass slower and large molecules will be excluded. Besides filtering, this method discriminates the used microbeads into k levels and thereby introduces a geometric multiplexity. Additionally, the exclusion of large entities enables the simultaneous detection of two target molecules, which exhibit the same affinity interaction. The hydrogel is formed through the combination of two macromonomers. One component is a homobifunctional polyethylene glycol linker, carrying a strained alkyne (PEG-BCN) and the second component is the azide-functionalized dendritic polyglycerol (dPG-N3). They react via the bioorthogonal strain-promoted azide alkyne cycloaddition (SPAAC). The hydrogel creates a solution-like environment for the diffusion of the investigated biomolecules all the while providing a stable, bioinert, and surface bound network.

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Section: Methodsmentioning

confidence: 99%

Section: ■ Experimental Sectionmentioning

confidence: 99%

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Spatial Separation of Microbeads into Detection Levels by a Bioorthogonal Porous Hydrogel for Size-Selective Analysis and Increased Multiplexity

et al. 2019

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“…It may contain a magnetic field to immobilize magnetic microspheres in a layer for optical stability so that a CCD camera instead of lasers can be used and more beads at once can be analyzed [ 145 , 148 ]. An example of a direct antigen multiplexing assay using a CCD camera is the typing and classification of O26, O55, O78, O118, O124, O127, O128, O142, O145, and O157 antigens on pathogenic Escherichia coli [ 154 ]. Mutatis mutandis , bead arrays are also used in a planar format in so-called “-omics” technologies enabling thousands of tests simultaneously while each of these tests analyzes a range of molecules in parallel [ 149 ].…”

Section: Analytical Microbiologymentioning

confidence: 99%

Modernization of Control of Pathogenic Micro-Organisms in the Food-Chain Requires a Durable Role for Immunoaffinity-Based Detection Methodology—A Review

Bergwerff

Debast

2021

Foods

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Food microbiology is deluged by a vastly growing plethora of analytical methods. This review endeavors to color the context into which methodology has to fit and underlines the importance of sampling and sample treatment. The context is that the highest risk of food contamination is through the animal and human fecal route with a majority of foodborne infections originating from sources in mass and domestic kitchens at the end of the food-chain. Containment requires easy-to-use, failsafe, single-use tests giving an overall risk score in situ. Conversely, progressive food-safety systems are relying increasingly on early assessment of batches and groups involving risk-based sampling, monitoring environment and herd/flock health status, and (historic) food-chain information. Accordingly, responsible field laboratories prefer specificity, multi-analyte, and high-throughput procedures. Under certain etiological and epidemiological circumstances, indirect antigen immunoaffinity assays outperform the diagnostic sensitivity and diagnostic specificity of e.g., nucleic acid sequence-based assays. The current bulk of testing involves therefore ante- and post-mortem probing of humoral response to several pathogens. In this review, the inclusion of immunoglobulins against additional invasive micro-organisms indicating the level of hygiene and ergo public health risks in tests is advocated. Immunomagnetic separation, immunochromatography, immunosensor, microsphere array, lab-on-a-chip/disc platforms increasingly in combination with nanotechnologies, are discussed. The heuristic development of portable and ambulant microfluidic devices is intriguing and promising. Tant pis, many new platforms seem unattainable as the industry standard. Comparability of results with those of reference methods hinders the implementation of new technologies. Whatever the scientific and technological excellence and incentives, the decision-maker determines this implementation after weighing mainly costs and business risks.

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“…It is a very versatile platform which uses standard equipment and consumables, and can be integrated with heating and cooling units, allowing it to be easily adaptable to the user’s needs. This technique has been successfully applied in bioanalysis such as detection of nucleic acids 32 , autoimmune diagnostics 33 , epitope mapping 34 , and serotyping of E. coli 35 as well as quantifying carboxyl groups on a microbead surface 36 .…”

Section: Introductionmentioning

confidence: 99%

A new reporter design based on DNA origami nanostructures for quantification of short oligonucleotides using microbeads

Choi

Schmidt

Tinnefeld

et al. 2019

Sci Rep

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The DNA origami technique has great potential for the development of brighter and more sensitive reporters for fluorescence based detection schemes such as a microbead-based assay in diagnostic applications. The nanostructures can be programmed to include multiple dye molecules to enhance the measured signal as well as multiple probe strands to increase the binding strength of the target oligonucleotide to these nanostructures. Here we present a proof-of-concept study to quantify short oligonucleotides by developing a novel DNA origami based reporter system, combined with planar microbead assays. Analysis of the assays using the VideoScan digital imaging platform showed DNA origami to be a more suitable reporter candidate for quantification of the target oligonucleotides at lower concentrations than a conventional reporter that consists of one dye molecule attached to a single stranded DNA. Efforts have been made to conduct multiplexed analysis of different targets as well as to enhance fluorescence signals obtained from the reporters. We therefore believe that the quantification of short oligonucleotides that exist in low copy numbers is achieved in a better way with the DNA origami nanostructures as reporters.

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

Cited by 8 publications

References 35 publications

Spatial Separation of Microbeads into Detection Levels by a Bioorthogonal Porous Hydrogel for Size-Selective Analysis and Increased Multiplexity

Spatial Separation of Microbeads into Detection Levels by a Bioorthogonal Porous Hydrogel for Size-Selective Analysis and Increased Multiplexity

Modernization of Control of Pathogenic Micro-Organisms in the Food-Chain Requires a Durable Role for Immunoaffinity-Based Detection Methodology—A Review

A new reporter design based on DNA origami nanostructures for quantification of short oligonucleotides using microbeads

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