Oriented T4 Bacteriophage Immobilization for E- coli Bacteria as A Potential Bioterrorism Agent Detection Based on Surface Plasmon Resonance Method

Document Type : Original Research

Authors

1 Nanobiotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

2 Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

3 Department of Biology, Roudehen Branch, Islamic Azad University, Roudehen, Iran

Abstract

Background and Aim: As a potential biological weapon, Escherichia coli (E. coli) is one of the most dangerous types of bacteria used in bioterrorism, which is responsible for many foodborne illnesses. In this study, a surface plasmon resonance biosensor (SPR) was designed to detect E. coli using oriented immobilization of T4 bacteriophage.
Methods: The surface of the gold disc was first modified with streptavidin. Then, the T4 bacteriophage was biotinylated. Optimization of biotin levels was performed using bacteriophage activity. After confirmation of bacteriophage biotinylation using the dynamic light scattering (DLS) method, T4 bacteriophage was oriented immobilized on the gold disk through the interactions between streptavidin and biotin.
Results: The equilibrium dissociation constant (KD) was found to be 10-17, which showed the great affinity of bacteriophage to interact with E. coli. Using the designed biosensor, E. coli was detected in the concentration range of 101 to 109 bacteria per ml.
Conclusion: Label-free and fast detection of E. coli are the advantages of the proposed biosensor.

Keywords


References 1. Tumbarski YD. Foodborne zoonotic agents and their food bioterrorism potential: A review. Bulg J Vet Med. 2020;23(2):147–59. 2. Zhang X, Shi C, Liu Z, Pan F, Meng R, Bu X, et al. Antibacterial activity and mode of action of ε-polylysine against escherichia coli O157:H7. J Med Microbiol. 2018;67(6):838–45. 3. Laster BA, Harris KB, Lucia LM, Castillo A, Savell JW. Efficacy of trimming chilled beef during fabrication to control Escherichia coli O157:H7 surrogates on subsequent subprimals. Meat Sci [Internet]. 2012;90(2):420–5. Available from: http://dx.doi.org/10.1016/j.meatsci.2011.08.011 4. Torun Ö, Hakki Boyaci I, Temür E, Tamer U. Comparison of sensing strategies in SPR biosensor for rapid and sensitive enumeration of bacteria. Biosens Bioelectron. 2012;37(1):53–60. 5. Homola J, Yee SS, Gauglitz G. Surface plasmon resonance sensors: review. Sensors Actuators, B Chem. 1999;54(1):3–15. 6. Tassa C, Duffner JL, Lewis TA, Weissleder R, Schreiber SL, Koehler AN, et al. Binding affinity and kinetic analysis of targeted small molecule-modified nanoparticles. Bioconjug Chem. 2010;21(1):14–9. 7. Morales-Morales HA, Vidal G, Olszewski J, Rock CM, Dasgupta D, Oshima KH, et al. Optimization of a reusable hollow-fiber ultrafilter for simultaneous concentration of enteric bacteria, protozoa, and viruses from water. Appl Environ Microbiol. 2003;69(7):4098–102. 8. Tolba M, Minikh O, Brovko LY, Evoy S, Griffiths MW. Oriented immobilization of bacteriophages for biosensor applications. Appl Environ Microbiol. 2010;76(2):528–35. 9. Favrin SJ, Jassim SA, Griffiths MW. Development and optimization of a novel immunomagnetic separation-bacteriophage assay for detection of Salmonella enterica serovar enteritidis in broth. Appl Environ Microbiol. 2001;67(1):217–24. 10. Gervais L, Gel M, Allain B, Tolba M, Brovko L, Zourob M, et al. Immobilization of biotinylated bacteriophages on biosensor surfaces. Sensors Actuators, B Chem. 2007;125(2):615–21. 11. Pantucek R, Doskar J, Ruzickova V, Kasparek P, Oracova E, Kvardova V, et al. Identification of bacteriophage types and their carriage in Staphylococcus aureus. Arch Virol. 2004;149:1689–703. 12. Kahánková J, Spanová A, Pantucek R, Horak D, Doskar J, Rittich B. Extraction of PCR-ready DNA from Staphylococcus aureus bacteriophages using carboxyl functionalized magnetic nonporous microspheres. J Chromatogr B. 2009;877:599–602. 13. Zaveta K, Lancok A, Marysko M, Pollert E. Superparamagnetic properties of γ-Fe2O3 particles: M¨ossbauer spectroscopy and d.c. magnetic measurements. Czechoslov J Phys. 2006;56:83–91. 14. Barrangou R, Yoon S, Breidt F, Fleming HP, Klaenhammer TR. Characterization of Six Leuconostoc fallax Bacteriophages Isolated from an Industrial Sauerkraut Fermentation †. Appl Environ Microbiol. 2002;68(11):5452–8. 15. Saberi F, Kamali M, Taheri RA, Ramandi MF, Mirnejad R. Development of Surface Plasmon Resonance-Based Immunosensor for Detection of. J Braz Chem Soc. 2016;27(11):1960–5. 16. Feng Y, Chen C, Su L, Hu S, Yu J, Chiu C. Evolution and pathogenesis of Staphylococcus aureus : lessons learned from genotyping and comparative genomics. 2007; 17. Bae T, Baba T, Hiramatsu K, Schneewind O. Prophages of Staphylococcus aureus Newman and their contribution to virulence. 2006;