Physical Biochemistry, University of Potsdam, 14476 Potsdam, Germany.
Institut Pasteur, Unité de Chimie des Biomolécules, 28 rue du Roux, 75015 Paris, France.
Viruses. 2018 Aug 15;10(8):431. doi: 10.3390/v10080431.
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 (), a highly contagious pathogen causing dysentery. We found that Sf6TSP not only bound O-antigen of 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-tagII 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 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 detection system.
由于抗生素耐药性的增加，噬菌体研究变得越来越重要。然而，为了进行噬菌体治疗，诊断技术必须得到改进。噬菌体携带黏附蛋白，这些蛋白可以与细菌细胞表面结合，例如尾丝蛋白（TSP），用于特定识别细菌 O-抗原多糖。TSP 是高度稳定的蛋白，因此可能适合整合到诊断工具中。我们使用噬菌体 Sf6 的 TSP 建立了两种应用，用于检测（），一种高度传染性的病原体，会导致痢疾。我们发现 Sf6TSP 不仅结合了血清型 Y 的 O-抗原，还结合了血清型 2a 的葡萄糖化 O-抗原。此外，质谱聚糖分析表明 Sf6TSP 可以耐受这些 O-抗原上的各种 O-乙酰化修饰。我们使用经过 Strep-tagII 修饰的 Sf6TSP 建立了基于微量滴定板的 ELISA 样尾丝吸附测定法（ELITA）。作为敏感筛选的替代方法，我们通过偶联到环境敏感染料来制备荧光标记的 Sf6TSP。该探针与 O-抗原 Y 的结合引起荧光强度增加 80%，发射最大值在可见光范围内。因此，Sf6TSP 探针为基于噬菌体 TSP 的高度特异性和敏感的 检测系统提供了一条有前途的途径。
