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Viruses 2018, 10(8), 431; https://doi.org/10.3390/v10080431

Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens

1
Physical Biochemistry, University of Potsdam, 14476 Potsdam, Germany
2
Institut Pasteur, Unité de Chimie des Biomolécules, 28 rue du Roux, 75015 Paris, France
3
CNRS UMR 3523, Institut Pasteur, 75015 Paris, France
4
National Reference Centre for Salmonella and other Bacterial Enterics, Robert Koch Institute, 38855 Wernigerode, Germany
Current address: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK.
*
Author to whom correspondence should be addressed.
Received: 29 May 2018 / Revised: 26 July 2018 / Accepted: 9 August 2018 / Published: 15 August 2018
(This article belongs to the Special Issue Biotechnological Applications of Phage and Phage-Derived Proteins)
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Abstract

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. View Full-Text
Keywords: Shigella flexneri; bacteriophage; tailspike proteins; O-antigen; serotyping; microtiter plate assay; fluorescence sensor Shigella flexneri; bacteriophage; tailspike proteins; O-antigen; serotyping; microtiter plate assay; fluorescence sensor
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Kunstmann, S.; Scheidt, T.; Buchwald, S.; Helm, A.; Mulard, L.A.; Fruth, A.; Barbirz, S. Bacteriophage Sf6 Tailspike Protein for Detection of Shigella flexneri Pathogens. Viruses 2018, 10, 431.

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