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Lectin-based detection of Escherichia coli and Staphylococcus aureus by flow cytometry

  • Olga D. HendricksonEmail author
  • Vadim D. Nikitushkin
  • Anatoly V. Zherdev
  • Boris B. Dzantiev
Original Paper
  • 104 Downloads

Abstract

This study develops a flow cytometry analysis of the bacterial pathogens Escherichia coli and Staphylococcus aureus based on a ligand–bioreceptor interaction. We used fluorescently labeled plant lectins as natural receptors that could specifically interact with the cell wall carbohydrates of bacteria. An epifluorescence microscopy was used as an additional approach to confirm and visualize lectin–carbohydrate interactions. The binding specificity of plant lectins to E. coli and S. aureus cells was studied, and wheat germ agglutinin, which provided high-affinity interactions, was selected as a receptor. Using this method, bacterial pathogens can be detected in concentrations of up to 106 cells/mL within 5 min. Their accessibility and universality make lectin reagents a promising tool to control a wide range of bacterial pathogens.

Keywords

Lectins Carbohydrates Escherichia coli Staphylococcus aureus Flow cytometry Detection 

Notes

Acknowledgements

This study was supported by the Russian Science Foundation (Grant 14-14-01131) with the exception of the microscopic studies of Mycobacterium that were supported by the Russian Foundation for Basic Research (Grant 17-04-00564_a). The authors are grateful to Dr. V.N. Kopyltsov (Federal Research and Clinical Center of Physical–Chemical Medicine, Moscow, Russia) for providing S. aureus cells and Dr. T.A. Yagudin (A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, Russia) for providing E. coli TG1 cells.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

203_2018_1613_MOESM1_ESM.docx (64 kb)
Supplementary material 1 (DOCX 63 KB)

References

  1. Abbasian F, Ghafar-Zadeh E, Magierowski S (2018) Microbiological sensing technologies: a review. Bioengineering (Basel) 5:20CrossRefGoogle Scholar
  2. Alvarez-Barrientos A, Arroyo J, Canton R, Nombela C, Sanchez-Perez M (2000) Applications of flow cytometry to clinical microbiology. Clin Microbiol Rev 13:167–195CrossRefGoogle Scholar
  3. Bain R, Cronk R, Hossain R, Bonjour S, Onda K, Wright J et al (2014) Global assessment of exposure to faecal contamination through drinking water based on a systematic review. Trop Med Int Health 19:917–927CrossRefGoogle Scholar
  4. Brooks SA (2017) Lectin Histochemistry: Historical Perspectives, State of the Art, and the Future. In: Pellicciari C, Biggiogera M (eds) Histochemistry of single molecules. Methods in molecular biology, vol 1560. Humana Press, New YorkGoogle Scholar
  5. Buzatu DA, Moskal TJ, Williams AJ, Cooper WM, Mattes WB, Wilkes JG (2014) An integrated flow cytometry-based system for real-time, high sensitivity bacterial detection and identification. PLoS One 4:e94254CrossRefGoogle Scholar
  6. Dan X, Liu W, Ng TB (2016) Development and applications of lectins as biological tools in biomedical research. Med Res Rev 36:221–247CrossRefGoogle Scholar
  7. Davey HM (2002) Flow cytometric techniques for the detection of microorganisms. Methods Cell Sci 24:91–97CrossRefGoogle Scholar
  8. Dmitriev BA, Toukach FV, Holst O, Rietschel ET, Ehlers S (2004) Tertiary structure of Staphylococcus aureus cell wall murein. J Bacteriol 186:7141–7148CrossRefGoogle Scholar
  9. Faria-Ramos I, Costa-de-Oliveira S, Barbosa J, Cardoso A, Santos-Antunes J, Rodrigues AG et al (2012) Detection of Legionella pneumophila on clinical samples and susceptibility assessment by flow cytometry. Eur J Clin Microbiol Infect Dis 31:3351–3357CrossRefGoogle Scholar
  10. Fernandez-Lago L, Vallejo FJ, Trujillano I, Vizcaino N (2000) Fluorescent whole-cell hybridization with 16S rRNA-targeted oligonucleotide probes to identify Brucella spp. by flow cytometry. J Clin Microbiol 38:2768–2771PubMedPubMedCentralGoogle Scholar
  11. He X, Zhou L, He D, Wang K, Cao J (2011) Rapid and ultrasensitive E. coli O157:H7 quantitation by combination of ligand magnetic nanoparticles enrichment with fluorescent nanoparticles based two-color flow cytometry. Analyst 136:4183–4191CrossRefGoogle Scholar
  12. Hendon-Dunn CL, Thomas SR, Taylor SC, Bacon J (2018) A flow cytometry method for assessing M. tuberculosis responses to antibiotics. Methods Mol Biol 1736:51–57CrossRefGoogle Scholar
  13. Hendrickson OD, Zherdev AV (2018) Analytical application of lectins. Crit Rev Anal Chem 48:279–292CrossRefGoogle Scholar
  14. Hendrickson OD, Smirnova NI, Zherdev AV, Gasparyan VK, Dzantiev BB (2017) Enzyme-linked lectinosorbent assay of Escherichia coli and Staphylococcus aureus. Appl Biochem Microbiol 53:107–113CrossRefGoogle Scholar
  15. Hirabayashi J (2014a) Lectin-based glycomics: how and when was the technology born? Methods Mol Biol 1200:225–242CrossRefGoogle Scholar
  16. Hirabayashi EJ (2014b) Lectins, methods and protocols. Humana Press, New YorkGoogle Scholar
  17. Holm C, Jespersen L (2003) A flow-cytometric Gram-staining technique for milk-associated bacteria. Appl Environ Microbiol 69:2857–2863CrossRefGoogle Scholar
  18. Holm C, Mathiasen T, Jespersen L (2004) A flow cytometric technique for quantification and differentiation of bacteria in bulk tank milk. J Appl Microbiol 97:935–941CrossRefGoogle Scholar
  19. Holtje JV (1998) Growth of the stress-bearing and shape-maintaining murein sacculus of Escherichia coli. Microbiol Mol Biol Rev 62:181–203PubMedPubMedCentralGoogle Scholar
  20. Kennedy D, Wilkinson MG (2017) Application of flow cytometry to the detection of pathogenic bacteria. Curr Issues Mol Biol 23:21–38CrossRefGoogle Scholar
  21. Law JW, Ab Mutalib NS, Chan KG, Lee LH (2014) Rapid methods for the detection of foodborne bacterial pathogens: principles, applications, advantages and limitations. Front Microbiol 5:770PubMedGoogle Scholar
  22. Levon K, Yu B (2003) Development of multivalent macromolecular ligands for enhanced detection of biological targets. Macromol Symp 201:111–118CrossRefGoogle Scholar
  23. McCarthy M, Culloty SC (2011) Optimization of two immunofluorescent antibodies for the detection of Escherichia coli using immunofluorescent microscopy and flow cytometry. Curr Mirobiol 62:402–408CrossRefGoogle Scholar
  24. Mudronova D (2015) Flow cytometry as an auxiliary tool for the selection of probiotic bacteria. Benef Microbes 6:727–734CrossRefGoogle Scholar
  25. Nagata Y, Burger MM (1974) Wheat germ agglutinin. Molecular characteristics and specificity for sugar binding. J Biol Chem 249:3116–3122PubMedGoogle Scholar
  26. Richter SG, Elli D, Kim HK, Hendrickx APA, Sorg JA, Schneewind O et al (2013) Small molecule inhibitor of lipoteichoic acid synthesis is an antibiotic for Gram-positive bacteria. Proc Nat Acad Sci 110:3531–3536CrossRefGoogle Scholar
  27. Rüger M, Bensch G, Tüngler R, Reichl U (2012) A flow cytometric method for viability assessment of Staphylococcus aureus and Burkholderia cepacia in mixed culture. Cytometry A 81:1055–1066CrossRefGoogle Scholar
  28. Rüger M, Ackermann M, Reichl U (2014) Species-specific viability analysis of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus in mixed culture by flow cytometry. BMC Microbiol 14:56CrossRefGoogle Scholar
  29. Seltman G, Holst O (2002) The bacterial cell wall. Springer, BerlinCrossRefGoogle Scholar
  30. Singhal N, Kumar M, Kanaujia PK, Virdi JS (2015) MALDI-TOF mass spectrometry: an emerging technology for microbial identification and diagnosis. Front Microbiol 6:791CrossRefGoogle Scholar
  31. Vouga M, Greub G (2016) Emerging bacterial pathogens: the past and beyond. Clin Microbiol Infect 22:12–21CrossRefGoogle Scholar
  32. Xue Y, Wilkes JG, Moskal TJ, Williams AJ, Cooper WM, Buzatu DA (2017) Flow-cytometry-based method to detect Escherichia coli and Shigella spp. using 16S rRNA-based probe. Curr Protoc Toxicol 71:2251–2258CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Olga D. Hendrickson
    • 1
    Email author
  • Vadim D. Nikitushkin
    • 1
  • Anatoly V. Zherdev
    • 1
  • Boris B. Dzantiev
    • 1
  1. 1.A.N. Bach Institute of BiochemistryFederal Research Center “Fundamentals of Biotechnology” of the Russian Academy of SciencesMoscowRussia

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