TY - GEN A1 - Kunstmann, Ruth Sonja A1 - Scheidt, Tom A1 - Buchwald, Saskia A1 - Helm, Alexandra A1 - Mulard, Laurence A. A1 - Fruth, Angelika A1 - Barbirz, Stefanie T1 - Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens T2 - Viruses N2 - Bacteriophage research is gaining more importance due to increasing antibiotic resistance. However, for treatment with bacteriophages, diagnostics have to be improved. Bacteriophages carry adhesion proteins, which bind to the bacterial cell surface, for example tailspike proteins (TSP) for specific recognition of bacterial O-antigen polysaccharide. TSP are highly stable proteins and thus might be suitable components for the integration into diagnostic tools. We used the TSP of bacteriophage Sf6 to establish two applications for detecting Shigella flexneri (S. flexneri), a highly contagious pathogen causing dysentery. We found that Sf6TSP not only bound O-antigen of S. flexneri serotype Y, but also the glucosylated O-antigen of serotype 2a. Moreover, mass spectrometry glycan analyses showed that Sf6TSP tolerated various O-acetyl modifications on these O-antigens. We established a microtiter plate-based ELISA like tailspike adsorption assay (ELITA) using a Strep-tag®II modified Sf6TSP. As sensitive screening alternative we produced a fluorescently labeled Sf6TSP via coupling to an environment sensitive dye. Binding of this probe to the S. flexneri O-antigen Y elicited a fluorescence intensity increase of 80% with an emission maximum in the visible light range. The Sf6TSP probes thus offer a promising route to a highly specific and sensitive bacteriophage TSP-based Shigella detection system. T3 - Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe - 472 KW - Shigella flexneri KW - bacteriophage KW - tailspike proteins KW - O-antigen KW - serotyping KW - microtiter plate assay KW - fluorescence sensor Y1 - 2018 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-417831 ER - TY - JOUR A1 - Kunstmann, Ruth Sonja A1 - Scheidt, Tom A1 - Buchwald, Saskia A1 - Helm, Alexandra A1 - Mulard, Laurence A. A1 - Fruth, Angelika A1 - Barbirz, Stefanie T1 - Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens JF - Viruses N2 - Bacteriophage research is gaining more importance due to increasing antibiotic resistance. However, for treatment with bacteriophages, diagnostics have to be improved. Bacteriophages carry adhesion proteins, which bind to the bacterial cell surface, for example tailspike proteins (TSP) for specific recognition of bacterial O-antigen polysaccharide. TSP are highly stable proteins and thus might be suitable components for the integration into diagnostic tools. We used the TSP of bacteriophage Sf6 to establish two applications for detecting Shigella flexneri (S. flexneri), a highly contagious pathogen causing dysentery. We found that Sf6TSP not only bound O-antigen of S. flexneri serotype Y, but also the glucosylated O-antigen of serotype 2a. Moreover, mass spectrometry glycan analyses showed that Sf6TSP tolerated various O-acetyl modifications on these O-antigens. We established a microtiter plate-based ELISA like tailspike adsorption assay (ELITA) using a Strep-tag®II modified Sf6TSP. As sensitive screening alternative we produced a fluorescently labeled Sf6TSP via coupling to an environment sensitive dye. Binding of this probe to the S. flexneri O-antigen Y elicited a fluorescence intensity increase of 80% with an emission maximum in the visible light range. The Sf6TSP probes thus offer a promising route to a highly specific and sensitive bacteriophage TSP-based Shigella detection system. KW - Shigella flexneri KW - bacteriophage KW - tailspike proteins KW - O-antigen KW - serotyping KW - microtiter plate assay KW - fluorescence sensor Y1 - 2018 U6 - https://doi.org/10.3390/v10080431 SN - 1999-4915 VL - 10 IS - 8 SP - 1 EP - 18 PB - Molecular Diversity Preservation International (MDPI) CY - Basel ER -