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Schiffer D.,Acib Austrian Center of Industrial Biotechnology | Tegl G.,University of Natural Resources and Life Sciences, Vienna | Vielnascher R.,University of Natural Resources and Life Sciences, Vienna | Weber H.,University of Graz | And 6 more authors.
Materials | Year: 2015

There is a strong need for simple and fast methods for wound infection determination. Myeloperoxidase, an immune system-derived enzyme was found to be a suitable biomarker for wound infection. Hence, alkoxysilane-derivatized Fast Blue RR was immobilized via simple hydrolytic polymerization. The resulting enzyme-responsive siloxane layers were incubated with myeloperoxidase, wound fluid or hemoglobin. The reaction was monitored via HPLC measurements and the color development quantified spectrophotometrically. Myeloperoxidase was indeed able to oxidize immobilized Fast Blue RR leading to a blue colored product. No conversion was detected in non-infected wound fluids. The visible color changes of these novel materials towards blue enable an easy distinction between infected and non-infected wound fluids. © 2015 by the authors.


Tegl G.,University of Natural Resources and Life Sciences, Vienna | Schiffer D.,University of Natural Resources and Life Sciences, Vienna | Schiffer D.,Austrian Center of Industrial Biotechnology | Sigl E.,Qualizyme GmbH | And 4 more authors.
Applied Microbiology and Biotechnology | Year: 2015

Wound infection is a severe complication causing delayed healing and risks for patients. Conventional methods of diagnosis for infection involve error-prone clinical description of the wound and time-consuming microbiological tests. More reliable alternatives are still rare, except for invasive and unaffordable gold standard methods. This review discusses the diversity of new approaches for wound infection determination. There has been progress in the detection methods of microorganisms, including the assessment of the diversity of the bacterial community present in a wound, as well as in the elaboration of specific markers. Another interesting strategy involves the quantification of enzyme activities in the wound fluid secreted by the immune system as response to infection. Color-changing substrates for these enzymes consequently have been shown to allow detection of an infection in wounds in a fast and easy way. Promising results were also delivered in measuring pH changes or detecting enhanced amounts of volatile molecules in case of infection. A simple and effective infection detection tool is not yet on the market, but innovative ideas pave the way for the investigation of fast and easy point-of-care devices. © 2015, Springer-Verlag Berlin Heidelberg.


Schiffer D.,Austrian Center of Industrial Biotechnology | Schiffer D.,University of Natural Resources and Life Sciences, Vienna | Tegl G.,University of Natural Resources and Life Sciences, Vienna | Heinzle A.,Austrian Center of Industrial Biotechnology | And 6 more authors.
Expert Review of Molecular Diagnostics | Year: 2015

There is a pressing need for point-of-care diagnostics indicating early stages of infection. Polymers can respond to enzymes secreted by microorganisms or released by the human immune system. This provokes either a direct color reaction or release of dyes, allowing early-stage detection of wound infections and contamination of medical devices. Conventional methods for the detection of infection indicators are based on slow, laboratory-based procedures and, consequently, do not allow a timely assessment. In contrast, polymer-based materials offer real-time responses in point-of-care devices that, in turn, allow therapists to amend treatment before the infection has become firmly established. The use of protein, polysaccharide and mixed polymer systems provides a sensitive means to detect the low levels of proteases and glycosyl hydrolases produced on initiation of infection in the clinical setting. These polymers can be easily fabricated into various forms that can be directly applied in diagnostic devices. © 2015 © Informa UK, Ltd.


PubMed | University of Vienna, University of Graz, Acib Austrian Center of Industrial Biotechnology and Qualizyme GmbH
Type: Journal Article | Journal: Biotechnology and bioengineering | Year: 2016

There is a strong need for simple and fast diagnostic tools for the detection of wound infection. Immune system-derived enzymes like myeloperoxidase are efficient biomarkers for wound infection that emerge in the early stage infection process. In this study, 5-amino-2-methoxyphenol was functionalized with alkoxysilane to allow visual detection of MPO on carrier materials, for example, in test strips. Indeed, MPO activity was visually detectable in short time in wound background. Oxidation of the substrate was followed spectrophotometrically and proved via HPLC. LC-ESI TOF and NMR analyses unveiled the reaction mechanism and a dimeric reaction product responsible for the visualization of MPO activity. The substrate specificity and sensitivity toward MPO detection was proved and tests with infected wound fluids were successfully performed. The study demonstrates the suitability of the novel MPO substrate for the detection of wound infection and the covalent immobilization on diagnostic carrier materials. Biotechnol. Bioeng. 2016;113: 2553-2560. 2016 Wiley Periodicals, Inc.


Tegl G.,University of Vienna | Rollett A.,University of Vienna | Dopplinger J.,University of Vienna | Gamerith C.,Qualizyme GmbH | And 3 more authors.
Carbohydrate Polymers | Year: 2016

There is a strong need of point-of-care diagnostics for early detection of wound infection. In this study, substrates based on functionalized chitosan were developed for visual detection of elevated lysozyme activity, an infection biomarker in wound fluids. For efficient hydrolysis by lysozyme, N-acetyl chitosan with a final degree of acetylation of around 50% was synthesized. N-acetylated chitosan and a chitosan-starch composite were labeled with structurally different dyes resulting in lysozyme-responsive biomaterials. Incubation with lysozyme in buffer and artificial wound fluid lead to a release of colored hydrolysis products already after 2 h incubation. Tests in human wound fluid from infected wounds indicated a clear visual color change after 2.5 h compared to control samples. A higher degree of swelling of the chitosan/starch containing substrate led to faster hydrolysis by lysozyme. This study demonstrates the potential of the lysozyme-responsive materials for diagnosis of wound infection and provides different diagnostic substrates for potential incorporation in point-of-care devices. © 2016 Elsevier Ltd. All rights reserved.

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