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Modified Bacteriophage Tail Fiber Proteins for Labeling, Immobilization, Capture, and Detection of Bacteria.
Journal article

Modified Bacteriophage Tail Fiber Proteins for Labeling, Immobilization, Capture, and Detection of Bacteria.

  • Dunne M Laboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zurich, Zurich, Switzerland.
  • Loessner MJ Laboratory of Food Microbiology, Institute of Food, Nutrition and Health, ETH Zurich, Zurich, Switzerland. martin.loessner@ethz.ch.
  • 2018-12-24
Published in:
  • Methods in molecular biology (Clifton, N.J.). - 2019
Bacteriophage
Foodborne bacteria
Long tail fiber
Pathogen detection
Phage S16
Rapid detection assay
Receptor-binding proteins
Salmonella
Bacteria
Bacterial Infections
Bacteriophages
Biotinylation
Immunoassay
Plasmids
Recombinant Proteins
Salmonella Phages
Viral Proteins
English A critical component of bacterial detection assays is choosing a suitable affinity molecule that retains sensitivity and specificity for the target pathogen over a wide range of in situ applications. Bacteriophages (phages) are bacterial viruses that bind and infect their host cells with unmatched specificity. Phage host range is often determined by their long tail fibers (LTFs) that mediate adsorption of the virus particle to potential bacterial host cells, by binding to specific cell surface receptors. The inherent specificity of the LTFs for distinct bacterial species makes them ideal candidates for development into recombinant affinity molecules. In this chapter, we describe the development of the Salmonella phage S16 LTF (S16 LTF) into an affinity molecule as part of a novel assay to detect Salmonella cells. The enzyme-linked long tail fiber assay (ELLTA) involves two steps: (1) Immobilization and separation of Salmonella cells using S16 LTF-coated paramagnetic beads (LTF-MBs), and (2) Labeling of bead-captured Salmonella using horseradish peroxidase-conjugated S16 LTF (HRP-LTF). Rapid HRP-mediated conversion of a chromogenic substrate provides visual confirmation for the presence of Salmonella. Overall, the ELLTA assay requires as little as 2 h to detect as few as 102 cfu/ml Salmonella cells from liquid culture. The absorbance of the enzyme-generated color substrate is largely proportional to the present bacterial concentrations between 102 and 107 cfu/ml, providing semiquantitative determination of Salmonella cell counts. The methodology described in this chapter can be adapted for other phage receptor-binding proteins, to develop ELLTAs for the detection of other relevant bacterial pathogens.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/52549
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