This study aims to detect pathogenic Escherichia coli (E. coli) bacteria using non-destructive fluorescence microscopy and micro-Raman spectroscopy.
Raman vibrational spectroscopy provides additional information regarding biochemical changes at the cellular level. We have used two nanomaterials zinc oxide nanoparticles (ZnO-NPs) and gold nanoparticles (Au-NPs) to detect pathogenic E. coli. The scanning electron microscope (SEM) with energy dispersive X-ray (EDAX) spectroscopy exhibit surface morphology and the elemental composition of the synthesized NPs. The metal NPs are useful contrast agents due to the surface plasmon resonance (SPR) to detect the signal intensity and hence the bacterial cells. The changes due to the interaction between cells and NPs are further correlated to the change in the surface charge and stiffness of the cell surface with the help of the fluorescence microscopic assay.
We conclude that when two E. coli strains (MTCC723 and MTCC443) and NPs are respectively mixed and kept overnight, the growth of bacteria are inhibited by ZnO-NPs due to changes in cell membrane permeability and intracellular metabolic system under fluorescence microscopy. However, SPR possessed Au-NPs result in enhanced fluorescence of both pathogens. In addition, with the help of Raman microscopy and element analysis, significant changes are observed when Au-NPs are added with the two strains as compared to ZnO-NPs due to protein, lipid and DNA/RNA induced conformational changes.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Altunbek M, Kuku G, Culha M (2016) Gold nanoparticles in single-cell analysis for surface enhanced Raman scattering (SERS). Molecules 21:1–18. https://doi.org/10.3390/molecules21121617
Ammendola S, Pasquali P, Pistoia C et al (2007) High-affinity Zn2+ uptake system ZnuABC is required for bacterial zinc homeostasis in intracellular environments and contributes to the virulence of Salmonella enterica. Infect Immun 75:5867–5876. https://doi.org/10.1128/IAI.00559-07
Arredondo AR, Dorval BL, Klibanov AM et al (2000) Rapid immune detection of Escherichia coli. Biotechnol Lett 22:547–550. https://doi.org/10.1023/A:1005616523897
Bardhan R, Grady NK, Cole JR et al (2009) Fluorescence enhancement by Au nanostructures: nanoshells and nanorods. ACS Nano 3:744–752. https://doi.org/10.1021/nn900001q
Bianchi MA, Del Rio D, Pellegrini N et al (2004) A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells. Lett Appl Microbiol 39:301–305. https://doi.org/10.1111/j.1472-765X.2004.01589.x
Chang YC, Yang CY, Sun RL et al (2013) Rapid single cell detection of Staphylococcus aureus by aptamer-conjugated gold nanoparticles. Sci Rep 39:1–7. https://doi.org/10.1038/srep01863
Dartnell LR, Roberts TA, Moore G et al (2013) Fluorescence characterization of clinically-important bacteria. PLoS ONE 8:1–13. https://doi.org/10.1371/journal.pone.0075270
Das S, Vasireddi R, Villa KH et al (2015) Enhancement mechanism of fluorescence intensity in presence of plasmonic nanoparticles. Proc SPIE 9792:28–33. https://doi.org/10.1117/12.2207565
Dobrindt U, Agerer F, Michaelis K et al (2003) Analysis of genome plasticity in pathogenic and commensal Escherichia coli isolates by use of DNA arrays. J Bacteriol 185:1831–1840. https://doi.org/10.1128/jb.185.6.1831-1840.2003
Fu Y, Zhang J, Lakowicz JR (2010) Plasmon-enhanced fluorescence from single fluorophores end-linked to gold nanorods. J Am Chem Soc 132:5540–5541. https://doi.org/10.1021/ja9096237
Galikowska E, Kunikowska D, Tokarska-Pietrzak E et al (2011) Specific detection of Salmonella enterica and Escherichia coli strains by using ELISA with bacteriophages as recognition agents. Eur J Clin Microbiol Infect Dis 30:1067–1073. https://doi.org/10.1007/s10096-011-1193-2
Gopinath A, Boriskina SV, Premasiri WR et al (2009) Plasmonic nanogalaxies: multiscale aperiodic arrays for surface-enhanced raman sensing. Nano Lett 9:3922–3929. https://doi.org/10.1021/nl902134r
Hamasha K, Mohaidat QI, Putnam RA et al (2013) Sensitive and specific discrimination of pathogenic and nonpathogenic Escherichia coli using Raman spectroscopy—a comparison of two multivariate analysis techniques. Biomed Opt Express 4:481–489. https://doi.org/10.1364/BOE.4.000481
Hameed A, Karthikeyan C, Ahamed A et al (2016) In vitro antibacterial activity of ZnO and Nd doped ZnO nanoparticles against ESBL producing Escherichia coli and Klebsiella pneumonia. Sci Rep 6:1–11. https://doi.org/10.1038/srep24312
He X, Patfield S, Hnasko R et al (2013) A polyclonal antibody based immunoassay detects seven subtypes of shiga toxin 2 produced by Escherichia coli in human and environmental samples. PLoS ONE 8:261–289. https://doi.org/10.1371/journal.pone.0076368
Järvinen AK, Laakso S, Piiparinen P et al (2009) Rapid identification of bacterial pathogens using a PCR- and microarray-based assay. BMC Microbiol 9:1–161. https://doi.org/10.1186/1471-2180-9-161
Kairyte K, Luksiene Z, Pucetaite M, Sablinskas V (2012) Differentiation of bacterial strains by means of surface enhanced FT-Raman spectroscopy. Lith J Phys 52:276–283. https://doi.org/10.3952/physics.v52i3.2480
Kang X, Li Y, Fan L et al (2012) Development of an ELISA-array for simultaneous detection of five encephalitis viruses. Virol J 9:1–8. https://doi.org/10.1186/1743-422X-9-56
Law JW, AbMutalib NS, Chan KG, Lee LH (2015) Rapid methods for the detection of foodborne bacterial pathogen: principles, applications, advantages and limitations. Front Microbiol 5:19. https://doi.org/10.3389/fmicb.2014.00770
Lazcka O, Del Campo FJ, Muñoz FX (2007) Pathogen detection: a perspective of traditional methods and biosensors. Biosens Bioelectron 22:1205–1217. https://doi.org/10.1016/j.bios.2006.06.036
Liu TY, Chen Y, Wang HH et al (2012) Differentiation of bacteria cell wall using Raman scattering enhanced by nanoparticle array. J Nanosci Nanotechnol 12:5004–5008. https://doi.org/10.1166/jnn.2012.4941
Maruyama T, Takata T et al (2003) Simple detection of point mutations in DNA oligonucleotides using SYBR Green I. Biotechnol Lett 25:1637–1641. https://doi.org/10.1023/a:1025661730518
Meder H, Baumstummler A, Chollet R et al (2012) Fluorescence-based rapid detection of microbiological contaminants in water samples. Sci World J 2012:10. https://doi.org/10.1100/2012/234858
Ming T, Zhao L, Yang Z et al (2009) Strong polarization dependence of plasmon enhanced fluorescence on single gold nanorods. Nano Lett 9:3896–3903. https://doi.org/10.1021/nl902095q
Mocan T, Matea CT, Pop T et al (2017) Development of nanoparticle-based optical sensors for pathogenic bacterial detection. J Nanobiotechnol 15:1–14. https://doi.org/10.1186/s12951-017-0260-y
Molina F, López-Acedo E, Tabla R et al (2015) Improved detection of Escherichia coli and coliform bacteria by multiplex PCR. BMC Biotechnol 15:2–9. https://doi.org/10.1186/s12896-015-0168-2
Nie S, Emory SR (1997) Probing Single Molecules and Single Nanoparticles by Surface-Enhanced Raman Scattering. Science 275:1102–1106. https://doi.org/10.1126/science.275.5303.1102
Nishi K, Isobe S, Zhu Y, Kiyama R (2015) Fluorescence-based bioassays for the detection and evaluation of food materials. Sensors 15:25831–25867. https://doi.org/10.3390/s151025831
Ou L, Chen Y, Su Y et al (2013) Application of silver nanoparticle-based SERS spectroscopy for DNA analysis in radiated nanopharyngeal carcinoma cells. J Raman Spectrosc 44:680–685. https://doi.org/10.1002/jrs.4269
Padmavathy N, Vijayaraghavan R (2011) Interaction of ZnO nanoparticles with microbes–a physio and biochemical assay. Biomed Nanotechnol 7:813–822. https://doi.org/10.1166/jbn.2011.1343
Queirós RB, de-los-Santos-Álvarez N, Noronha JP, Sales MGF (2013) A label-free DNA aptamer-based impedance biosensor for the detection of E. coli outer membrane proteins. Sensors Actuators B 181:766–772. https://doi.org/10.1016/j.snb.2013.01.062
Rasooly A, Herold KE (2008) Food microbial pathogen detection using DNA microarray technologies. Foodborne Pathog Dis 5:531–550. https://doi.org/10.1089/fpd.2008.0119
Stöckel S, Kirchhoff J, Neugebauer U et al (2016) The application of Raman spectroscopy for the detection and identification of microorganisms. J Raman Spectrosc 47(1):4789–5109. https://doi.org/10.1002/jrs.4844
Tam F, Goodrich GP, Johnson BR, Halas NJ (2007) Plasmonic enhancement of molecular fluorescence. Nano Lett 7:496–501. https://doi.org/10.1021/nl062901x
Teng L, Wang X, Wang X et al (2016) Label-free, rapid and quantitative phenotyping of stress response in Escherichia coli via ramanome. Sci Rep 6:34359. https://doi.org/10.1038/srep34359
Vasireddi R, Javvaji B, Vardhan H et al (2017) Growth of zinc oxide nanorod structures: pressure controlled hydrothermal process and growth mechanism. J Mater Sci 52:2007–2020. https://doi.org/10.1007/s10853-016-0489-0
Velusamy V, Arshak K, Korostynska O et al (2010) An overview of food borne pathogen detection: In the perspective of biosensors. Biotechnol Adv 28:232–254. https://doi.org/10.1016/j.biotechadv.2009.12.004
Wang L, Li Y, Chu J et al (2012) Development and application of a simple loop-mediated isothermal amplification method on rapid detection of Listeria monocytogenes strains. Mol Biol Rep 39:445–449. https://doi.org/10.1007/s11033-011-0757-7
Xia X, Meng J, McDermott PF et al (2010) Presence and characterization of shiga toxin-producing Escherichia coli and other potentially diarrheagenic E. coli strains in retail meats. Appl Environ Microbiol 76:1709–1717. https://doi.org/10.1128/AEM.01968-09
Xu JG, Cheng BK, Jing HQ (1999) Escherichia coli O157: H7 and Shiga-like-toxin producing Escherichia coli in China. World J Gastroenterol 5:191–194. https://doi.org/10.3748/wjg.v5.i3.191
Ye BC, Zhang M, Yin BC (2012) Nanomaterial-enhanced fluorescence polarization and its application. In: Ye BC, Zhang M, Yin BC (eds) Nano-bio probe design and its application for biochemical analysis. Springer, Berlin, pp 3–25
Zhao X, Li Y, Wang L, You L et al (2010a) Development and application of a loop-mediated isothermal amplification method on rapid detection Escherichia coli O157 strains from food samples. Mol Biol Rep 37:2183–2188. https://doi.org/10.1007/s11033-009-9700-6
Zhao X, Wang L, Chu J et al (2010b) Rapid detection of Vibrio parahaemolyticus strains and virulent factors by loop-mediated isothermal amplification assays. Food Sci Biotechnol 19:1191–1197. https://doi.org/10.1007/s10068-010-0170-3
Zhao X, Wang L, Chu J et al (2010c) Development and application of a rapid and simple loop-mediated isothermal amplification method for food-borne Salmonella detection. Food Sci Biotechnol 19:1655–1659. https://doi.org/10.1007/s10068-010-0234-4
Zhou H, Yang D, Mircescu NE et al (2015) Surface-enhanced Raman scattering detection of bacteria on microarrays at single cell levels using silver nanoparticles. Microchim Acta 182:2259–2266. https://doi.org/10.1007/s00604-015-1570-0
Zhu L, He J, Cao X, Huang K et al (2016) Development of a double antibody sandwich ELISA for rapid detection of Bacillus Cereus in food. Sci Rep 6:1–10. https://doi.org/10.1038/srep16092
The author Gargibala would like to thank for Prof Roy Mahapatra, iMEMS (Laboratory of Integrative Multiscale Engineering Materials and Systems), Prof Siva Umapathy, IPC (Inorganic Physical Chemistry), IISc, Bangalore and BIHER, Chennai management provided the permission and facility to do the above work. DRM and GS thankfully acknowledge funding support through project IDC-Water under the Indo-German Science and Technology (IGSTC) 2 + 2 program to carry out the research.
This work was funded by the Indo-German Science and Technology Centre (IGSTC), Foundation Grant IDC-Water/16/2017.
Conflict of interest
The authors declare no conflict of interests.
This article does not contain any studies with human participants or animals performed by any of the authors.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Satpathy, G., Chandra, G.K., Manikandan, E. et al. Pathogenic Escherichia coli (E. coli) detection through tuned nanoparticles enhancement study. Biotechnol Lett (2020). https://doi.org/10.1007/s10529-020-02835-y
- E. coli
- Gold and ZnO nanoparticles
- Fluorescence spectroscopy