Advertisement

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

  • Matthew Dunne
  • Martin J. LoessnerEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1918)

Abstract

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.

Key words

Bacteriophage Receptor-binding proteins Long tail fiber Salmonella Phage S16 Pathogen detection Foodborne bacteria Rapid detection assay 

Notes

Acknowledgments

We thank Jenna M. Denyes for her significant contribution toward the development of the ELLTA methodology and for performing the LTF-MB pulldown assays and preliminary HRP-LTF detection tests. We are also grateful for the continuous support and ideas received from Jochen Klumpp. We thank Roger Stephan and Herbert Hächler (University of Zurich, Zurich, Switzerland), and Herbert Schmidt and Agnes Weiss (Hohenheim University, Germany) for valuable discussions regarding Salmonella detection, and finally Stefan Miller (Regensburg, Germany) for advice regarding the production and use of recombinant phage proteins. The project was funded by the AiF/FEI, Bundesministerium für Wirtschaft und Technologie, Berlin, Germany (Grant number 16756 N).

References

  1. 1.
    Bell RL, Jarvis KG, Ottesen AR et al (2016) Recent and emerging innovations in Salmonella detection: a food and environmental perspective. Microb Biotechnol 9:279–292CrossRefGoogle Scholar
  2. 2.
    Cudjoe KS, Krona R, Olsen E (1994) IMS: a new selective enrichment technique for detection of Salmonella in foods. Int J Food Microbiol 23:159–165CrossRefGoogle Scholar
  3. 3.
    de Cássia dos Santos da Conceição R, Moreira ÂN, Ramos RJ et al (2008) Detection of Salmonella sp in chicken cuts using immunomagnetic separation. Braz J Microbiol 39:173–177CrossRefGoogle Scholar
  4. 4.
    Mansfield LP, Forsythe SJ (2000) The detection of Salmonella using a combined immunomagnetic separation and ELISA end-detection procedure. Lett Appl Microbiol 31:279–283CrossRefGoogle Scholar
  5. 5.
    Muldoon MT, Teaney G, Jingkun LI et al (2007) Bacteriophage-based enrichment coupled to Immunochromatographic strip–based detection for the determination of Salmonella in meat and poultry. J Food Prot 70:2235–2242CrossRefGoogle Scholar
  6. 6.
    Nilsson AS (2014) Phage therapy—constraints and possibilities. Ups J Med Sci 119:192–198CrossRefGoogle Scholar
  7. 7.
    Schooley RT, Biswas B, Gill JJ et al (2017) Development and use of personalized bacteriophage-based therapeutic cocktails to treat a patient with a disseminated resistant Acinetobacter baumannii infection. Antimicrob Agents Chemother 61(10):00954–00917CrossRefGoogle Scholar
  8. 8.
    Singh A, Poshtiban S, Evoy S (2013) Recent advances in bacteriophage based biosensors for food-borne pathogen detection. Sensors (Basel) 13:1763–1786CrossRefGoogle Scholar
  9. 9.
    Schmelcher M, Loessner MJ (2014) Application of bacteriophages for detection of foodborne pathogens. Bacteriophage 4:e28137CrossRefGoogle Scholar
  10. 10.
    Zinno P, Devirgiliis C, Ercolini D et al (2014) Bacteriophage P22 to challenge Salmonella in foods. Int J Food Microbiol 191:69–74CrossRefGoogle Scholar
  11. 11.
    Guenther S, Huwyler D, Richard S et al (2009) Virulent bacteriophage for efficient biocontrol of Listeria monocytogenes in ready-to-eat foods. Appl Environ Microbiol 75:93–100CrossRefGoogle Scholar
  12. 12.
    Guenther S, Herzig O, Fieseler L et al (2012) Biocontrol of Salmonella Typhimurium in RTE foods with the virulent bacteriophage FO1-E2. Int J Food Microbiol 154:66–72CrossRefGoogle Scholar
  13. 13.
    Kittler S, Fischer S, Abdulmawjood A et al (2013) Effect of bacteriophage application on campylobacter jejuni loads in commercial broiler flocks. Appl Environ Microbiol 79:7525–7533CrossRefGoogle Scholar
  14. 14.
    Borie C, Albala I, Sánchez P et al (2008) Bacteriophage treatment reduces Salmonella colonization of infected chickens. Avian Dis 52:64–67CrossRefGoogle Scholar
  15. 15.
    Kim JS, Hosseindoust A, Lee SH et al (2017) Bacteriophage cocktail and multi-strain probiotics in the feed for weanling pigs: effects on intestine morphology and targeted intestinal coliforms and Clostridium. Animal 11:45–53CrossRefGoogle Scholar
  16. 16.
    Kazi M, Annapure US (2016) Bacteriophage biocontrol of foodborne pathogens. J Food Sci Technol 53:1355–1362CrossRefGoogle Scholar
  17. 17.
    Klumpp J, Loessner MJ (2013) Listeria phages. Bacteriophage 3:e26861CrossRefGoogle Scholar
  18. 18.
    Balasubramanian S, Sorokulova IB, Vodyanoy VJ et al (2007) Lytic phage as a specific and selective probe for detection of Staphylococcus aureus—a surface plasmon resonance spectroscopic study. Biosens Bioelectron 22:948–955CrossRefGoogle Scholar
  19. 19.
    Lakshmanan RS, Guntupalli R, Hu J et al (2007) Phage immobilized magnetoelastic sensor for the detection of Salmonella typhimurium. J Microbiol Methods 71:55–60CrossRefGoogle Scholar
  20. 20.
    Lakshmanan RS, Guntupalli R, Hu J et al (2007) Detection of Salmonella typhimurium in fat free milk using a phage immobilized magnetoelastic sensor. Sensors Actuators B Chem 126:544–550CrossRefGoogle Scholar
  21. 21.
    Laube T, Cortés P, Llagostera M et al (2013) Phagomagnetic immunoassay for the rapid detection of Salmonella. Appl Microbiol Biotechnol 98:1795–1805CrossRefGoogle Scholar
  22. 22.
    Ackermann H-W (2007) 5500 Phages examined in the electron microscope. Arch Virol 152:227–243CrossRefGoogle Scholar
  23. 23.
    Fokine A, Rossmann MG (2014) Molecular architecture of tailed double-stranded DNA phages. Bacteriophage 4:e28281CrossRefGoogle Scholar
  24. 24.
    Leiman PG, Arisaka F, van RMJ et al (2010) Morphogenesis of the T4 tail and tail fibers. Virol J 7:355CrossRefGoogle Scholar
  25. 25.
    Spinelli S, Campanacci V, Blangy S et al (2006) Modular structure of THE receptor binding proteins of Lactococcus lactis Phages THE RBP STRUCTURE OF THE TEMPERATE PHAGE TP901-1. J Biol Chem 281:14256–14262CrossRefGoogle Scholar
  26. 26.
    Taylor NMI, Prokhorov NS, Guerrero-Ferreira RC et al (2016) Structure of the T4 baseplate and its function in triggering sheath contraction. Nature 533:346–352CrossRefGoogle Scholar
  27. 27.
    Trojet SN, Caumont-Sarcos A, Perrody E et al (2011) The gp38 Adhesins of the T4 superfamily: a complex modular determinant of the Phage’s host specificity. Genome Biol Evol 3:674–686CrossRefGoogle Scholar
  28. 28.
    Bartual SG, Garcia-Doval C, Alonso J et al (2010) Two-chaperone assisted soluble expression and purification of the bacteriophage T4 long tail fibre protein gp37. Protein Expr Purif 70:116–121CrossRefGoogle Scholar
  29. 29.
    Marti R, Zurfluh K, Hagens S et al (2013) Long tail fibres of the novel broad-host-range T-even bacteriophage S16 specifically recognize Salmonella OmpC. Mol Microbiol 87:818–834CrossRefGoogle Scholar
  30. 30.
    Singh A, Arutyunov D, McDermott MT et al (2011) Specific detection of campylobacter jejuni using the bacteriophage NCTC 12673 receptor binding protein as a probe. Analyst 136:4780–4786CrossRefGoogle Scholar
  31. 31.
    Denyes JM, Dunne M, Steiner S et al (2017) Modified bacteriophage S16 long tail fiber proteins for rapid and specific immobilization and detection of Salmonella cells. Appl Environ Microbiol 83(12):e00277–e00217CrossRefGoogle Scholar
  32. 32.
    Schmidt A, Rabsch W, Broeker NK et al (2016) Bacteriophage tailspike protein based assay to monitor phase variable glucosylations in Salmonella O-antigens. BMC Microbiol 16:207CrossRefGoogle Scholar
  33. 33.
    Waseh S, Hanifi-Moghaddam P, Coleman R et al (2010) Orally administered P22 phage tailspike protein reduces Salmonella colonization in chickens: prospects of a novel therapy against bacterial infections. PLoS One 5:e13904CrossRefGoogle Scholar
  34. 34.
    Singh A, Arya SK, Glass N et al (2010) Bacteriophage tailspike proteins as molecular probes for sensitive and selective bacterial detection. Biosens Bioelectron 26:131–138CrossRefGoogle Scholar
  35. 35.
    Javed MA, Poshtiban S, Arutyunov D et al (2013) Bacteriophage receptor binding protein based assays for the simultaneous detection of campylobacter jejuni and campylobacter coli. PLoS One 8:e69770CrossRefGoogle Scholar
  36. 36.
    Chua JE, Manning PA, Morona R (1999) The Shigella flexneri bacteriophage Sf6 tailspike protein (TSP)/endorhamnosidase is related to the bacteriophage P22 TSP and has a motif common to exo- and endoglycanases, and C-5 epimerases. Microbiology 145(7):1649–1659CrossRefGoogle Scholar
  37. 37.
    Steinbacher S, Baxa U, Miller S et al (1996) Crystal structure of phage P22 tailspike protein complexed with Salmonella sp. O-antigen receptors. Proc Natl Acad Sci U S A 93:10584–10588CrossRefGoogle Scholar
  38. 38.
    Singh A, Arutyunov D, Szymanski CM et al (2012) Bacteriophage based probes for pathogen detection. Analyst 137:3405–3421CrossRefGoogle Scholar
  39. 39.
    Tétart F, Desplats C, HM K (1998) Genome plasticity in the distal tail fiber locus of the T-even bacteriophage: recombination between conserved motifs swaps adhesin specificity1. J Mol Biol 282:543–556CrossRefGoogle Scholar
  40. 40.
    Riede I, Drexler K, Schwarz H et al (1987) T-even-type bacteriophages use an adhesin for recognition of cellular receptors. J Mol Biol 194:23–30CrossRefGoogle Scholar
  41. 41.
    Bartual SG, Otero JM, Garcia-Doval C et al (2010) Structure of the bacteriophage T4 long tail fiber receptor-binding tip. Proc Natl Acad Sci U S A 107:20287–20292CrossRefGoogle Scholar
  42. 42.
    Henning U, Jann K (1979) Two-component nature of bacteriophage T4 receptor activity in Escherichia coli K-12. J Bacteriol 137:664–666PubMedPubMedCentralGoogle Scholar
  43. 43.
    Yu S, Yu F, Liu L et al (2016) Which one of the two common reporter systems is more suitable for chemiluminescent enzyme immunoassay: alkaline phosphatase or horseradish peroxidase? Luminescence 31:888–892CrossRefGoogle Scholar
  44. 44.
    Marusich EI, Kurochkina LP, VV M (1998) Chaperones in bacteriophage T4 assembly. Biochemistry (Mosc) 63:399–406Google Scholar
  45. 45.
    Matsui T, Griniuviené B, Goldberg E et al (1997) Isolation and characterization of a molecular chaperone, gp57A, of bacteriophage T4. J Bacteriol 179:1846–1851CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Laboratory of Food Microbiology, Institute of Food, Nutrition and HealthETH ZurichZurichSwitzerland

Personalised recommendations