Fabrication of a Highly Sensitive Nanoaptasensor for Rapid and Direct Detection of Escherichia Coli (Nasec) in Drinking Water
DOI:
https://doi.org/10.29356/jmcs.v69i2.2146Keywords:
E. coli, aptamer, aptasensor, drinking waterAbstract
Abstract. In this work, a novel nanoaptasensor (NASEc) for rapid and direct detection of Escherichia coli (E. coli) in drinking water was designed and constructed. The sensor is based on electrical conductivity measurements between a set of bare gold electrodes and a set of modified gold electrodes. The modified electrode set was covered with an aptamer layer (AE10) to improve affinity for E. coli and better binding to the electrode surface. Using the modified nanostructured layer with multiwall carbon nanotubes (MWCNT), the high sensitivity of the sensor was achieved and allowed to detect the presence of E. coli in quantities of a single colony forming units (CFU) per 100 mL. This sensitivity meets the relevant international drinking water quality standards. Each step in the process of the NASEc fabrication was verified and proved by different analytical techniques at a nanometric and molecular level: UV-vis and Raman Spectroscopy, AFM, and SEM. This portable, simple, and reusable nanoaptasensor with high selectivity and affinity offers faster detection (3 min) in the presence of E. coli compared to conventional colony forming units (CFU) counting-based methods (24 to 48 h). NASEc is designed for direct measurements without any pretreatment of samples and is helpful in potable water within a conductivity range of 50 to 1300 µS/cm.
Resumen. En este trabajo, se diseñó y construyó un nuevo nanoaptasensor (NASEc) para la detección rápida y directa de Escherichia coli (E. coli) en agua potable. El sensor se basa en mediciones de conductividad eléctrica entre un conjunto de electrodos de oro sin modificar y un conjunto de electrodos de oro modificado. El conjunto de electrodos modificado se cubrió con una capa de aptámero (AE10) para mejorar la afinidad por E. coli y una mejor unión a la superficie del electrodo. Utilizando la capa nanoestructurada modificada con nanotubos de carbono de paredes múltiples (MNTC) se logró la alta sensibilidad del sensor y permitió detectar la presencia de E. coli en cantidades de una sola unidad formadora de colonias (UFC) por 100 ml. Esta sensibilidad cumple con los estándares internacionales pertinentes de calidad del agua potable. Cada paso en el proceso de fabricación de NASEc fue verificado y probado mediante diferentes técnicas analíticas a nivel nanométrico y molecular: espectroscopia UV-vis y Raman, AFM y SEM. Este nanoaptasensor portátil, simple y reutilizable con alta selectividad y afinidad ofrece una detección más rápida (3 min) en presencia de E. coli en comparación con los métodos convencionales basados en el recuento de unidades formadoras de colonias (UFC) (24 a 48 h). NASEc está diseñado para mediciones directas sin ningún tratamiento previo de muestras y es útil en agua potable dentro de un rango de conductividad de 50 a 1300 µS/cm.
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References
Zhao, Y. W.; Wang, H. X.; Jia, G. C.; Li, Z. Sensors. 2018, 18, 1–16. DOI: https://doi.org/10.3390/s18010318. DOI: https://doi.org/10.3390/s18010318
Rompré, A.; Servais, P.; Baudart, J.; De-Roubin, M. R.; Laurent, P. J. Microbiol. Methods. 2002, 49, 31–54. DOI: https://doi.org/10.1016/S0167-7012(01)00351-7. DOI: https://doi.org/10.1016/S0167-7012(01)00351-7
Khan, F. M.; Gupta, R.; Sekhri, S. Environ. Sci. Pollut. Res. 2021, 28, 60778-60786. DOI: https://doi.org/10.1007/s11356-021-14983-3. DOI: https://doi.org/10.1007/s11356-021-14983-3
Wu, W.; Li, M.; Wang, Y.; Ouyang, H.; Wang, L.; Li, C.; Cao, Y.; Meng, Q.; Lu, J. Nanoscale Res. Lett. 2012, 7, 658. DOI: https://doi.org/10.1186/1556-276x-7-658. DOI: https://doi.org/10.1186/1556-276X-7-658
Eaton, A. D.; Clesceri, L. S.; Rice, E. W.; Greenberg, A. E.; Franson, M. A. H., in: Standard Methods for the Examination of Water and Wastewater, 21st ed., American Public Health Association, Washington, DC, 2005.
Walker, D. I.; McQuillan, J.; Taiwo, M.; Parks, R.; Stenton, C. A.; Morgan, H.; Mowlem, M. C.; Lees, D. N. Water Res. 2017, 126, 101–110. DOI: https://doi.org/10.1016/j.watres.2017.08.032. DOI: https://doi.org/10.1016/j.watres.2017.08.032
Kuo, J. T.; Cheng, C. Y.; Huang, H. H.; Tsao, C. F.; Chung, Y. C. J. Ind. Microbiol. Biotechnol. 2010, 37, 237–244. DOI: https://doi.org/10.1007/s10295-009-0666-0. DOI: https://doi.org/10.1007/s10295-009-0666-0
Serra, B.; Morales, M. D.; Zhang, J.; Reviejo, A. J.; Hall, E. H.; Pingarron, J. M. Anal. Chem. 2005, 77, 8115–8121. DOI: https://doi.org/10.1021/ac051327r. DOI: https://doi.org/10.1021/ac051327r
Ionescu, R. E. Escherichia coli: Recent Adv. Physiol., Pathog. Biotechnol. Appl. 2017. DOI: https://doi.org/10.5772/67392. DOI: https://doi.org/10.5772/67392
Hesari, N.; Alum, A.; Elzein, M.; Abbaszadegan, M. Enzyme Microb. Technol. 2016, 83, 22–28. DOI: https://doi.org/10.1016/j.enzmictec.2015.11.007. DOI: https://doi.org/10.1016/j.enzmictec.2015.11.007
Jiang, Y. S.; Riedel, T. E.; Popoola, J. A.; Morrow, B. R.; Cai, S.; Ellington, A. D.; Bhadra, S. Water Res. 2018, 131, 186–195. DOI: https://doi.org/10.1016/j.watres.2017.12.023. DOI: https://doi.org/10.1016/j.watres.2017.12.023
Mallya, A. N.; Sowmya, P.; Ramamurthy, P. C., in: Organic Nanocomposite Sensor for Detection of Escherichia coli, 2014 IEEE 2nd International Conference on Emerging Electronics (ICEE), Bengaluru, India, 2014. DOI: https://doi.org/10.1109/ICEmElec.2014.7151202
Khang, J.; Kim, D.; Chung, K. W.; Lee, J. H. Talanta. 2016, 147, 177–183. DOI: https://doi.org/10.1016/j.talanta.2015.09.055
Van Poucke, S. O.; Nelis, H. Appl. Environ. Microbiol. 1995, 61, 4505–4509. DOI: https://doi.org/10.1128/aem.61.12.4505-4509.1995. DOI: https://doi.org/10.1128/aem.61.12.4505-4509.1995
Loaiza, Ó. A.; Campuzano, S.; Pedrero, M.; García, P.; Pingarrón, J. M. Analyst. 2009, 134, 34–37. DOI: https://doi.org/10.1039/b815307h. DOI: https://doi.org/10.1039/B815307H
Wutor, V. C.; Togo, C. A.; Limson, J. L.; Pletschke, B. I. Enzyme Microb. Technol. 2007, 40, 1512–1517. DOI: https://doi.org/10.1016/j.enzmictec.2006.10.039
Su, L.; Zhang, P.; Zheng, D.; Wang, Y.; Zhong, R. Optoelectron. Lett. 2015, 11, 157–160. DOI: https://doi.org/10.1007/s11801-015-4216-x
Gunda, N. S. K.; Dasgupta, S.; Mitra, S. K. PLOS ONE. 2017, DOI: https://doi.org/10.1371/journal.pone.0183234. DOI: https://doi.org/10.1371/journal.pone.0183234
Zuo, P.; Li, X.; Dominguez, D. C.; Ye, B. C. Lab Chip. 2013, 13, 3921–3928. DOI: https://doi.org/10.1039/c3lc50654a. DOI: https://doi.org/10.1039/c3lc50654a
Mitra, S. Anal. Methods. 2014, 6, 6236–6246. DOI: https://doi.org/10.1039/C4AY01245C
Frahm, E.; Obst, U. J. Microbiol. Methods. 2003, 52, 123–131. DOI: https://doi.org/10.1016/s0167-7012(02)00150-1. DOI: https://doi.org/10.1016/S0167-7012(02)00150-1
McCracken, K. E.; Angus, S. V.; Reynolds, K. A.; Yoon, J. Y. Sci. Rep. 2016, 6, 1–13. DOI: https://doi.org/10.1038/srep27529
Berg, J. D.; Fiksdal, L. Appl. Environ. Microbiol. 1988, 54, 2118–2122. DOI: https://doi.org/10.1128/aem.54.8.2118-2122.1988. DOI: https://doi.org/10.1128/aem.54.8.2118-2122.1988
Sun, H.; Choy, T. S.; Zhu, D. R.; Yam, W. C.; Fung, Y. S. Biosens. Bioelectron. 2009, 24, 1405–1410. DOI: https://doi.org/10.1016/j.bios.2008.08.008. DOI: https://doi.org/10.1016/j.bios.2008.08.008
Kim, Y. S.; Song, M. Y.; Jurng, J.; Kim, B. C. Anal. Biochem. 2013, 436, 22–28. DOI: https://doi.org/10.1016/j.ab.2013.01.014. DOI: https://doi.org/10.1016/j.ab.2013.01.014
Evtugyn, G.; Porfireva, A.; Ryabova, M.; Hianik, T. Electroanalysis. 2008, 20, 2310–2316. DOI: https://doi.org/10.1002/elan.200804345. DOI: https://doi.org/10.1002/elan.200804345
Téllez-Plancarte, A.; Haro-Poniatowski, E.; Piquart, M.; Morales-Méndez, J. G.; Lara-Cruz, C.; Jiménez-Salazar, J. E.; Damian-Matsumura, P.; Escobar-Alarcón, L.; Batina, N. Nanomaterials. 2018, 8, 549. DOI: https://doi.org/10.3390/nano8070549. DOI: https://doi.org/10.3390/nano8070549
Gómez-Reyes, A. R. E.; Monroy, O. A.; Batina, N.; Jiménez, D.; Abad, A. R.; Constantino, R.; Montero, D. P.; Tapia, F. O.; Ramírez, F.; Velázquez, E.; Morales, I.; Hernández, P.; Solís, A.; Hernández, J. de J. Mex. Pat., 2021.
Zhang, S.; Wang, X.; Li, T.; Liu, L.; Wu, H. C.; Luo, M.; Li, J. Langmuir. 2015, 31, 10094–10099. DOI: https://doi.org/10.1021/acs.langmuir.5b01272. DOI: https://doi.org/10.1021/acs.langmuir.5b01272
Ozkan-Ariksoysal, D.; Kayran, Y. U.; Yilmaz, F. F.; Ciucu, A. A.; David, I. G.; David, V.; Hosgor-Limoncu, M.; Ozsoz, M. Talanta. 2017, 166, 27–35. DOI: https://doi.org/10.1016/j.talanta.2017.01.005. DOI: https://doi.org/10.1016/j.talanta.2017.01.005
Yang, M.; Peng, Z.; Ning, Y.; Chen, Y.; Zhou, Q.; Deng, L. Sensors. 2013, 13, 6865–6881. DOI: https://doi.org/10.3390/s130506865. DOI: https://doi.org/10.3390/s130506865
Ocaña, C.; Pacios, M.; del Valle, M. Sensors. 2012, 12, 3037–3048. DOI: https://doi.org/10.3390/s120303037. DOI: https://doi.org/10.3390/s120303037
Murphy, H.; Papakonstantinou, P.; Okpalugo, T. I. T. J. Vac. Sci. Technol., B: Microelectron. Nanometer Struct. --Process., Meas., Phenom. 2006, 24, 715–720. DOI: https://doi.org/10.1116/1.2180257
Le, V. T.; Ngo, C. L.; Le, Q. T.; Ngo, T. T.; Nguyen, D. N.; Vu, M. T. Adv. Nat. Sci. Nanosci. Nanotechnol. 2013, 4, 4–9. DOI: https://doi.org/10.1088/2043-6262/4/3/035017
Maquelin, K.; Kirschner, C.; Choo-Smith, L.-P.; Ngo-Thi, N. A.; van Vreeswijk, T.; Stämmler, M.; Endtz, H. P.; Bruining, H. A.; Naumann, D.; Puppels, G. J. J. Clin. Microbiol. 2003, 41, 324–329. DOI: https://doi.org/10.1128/jcm.41.1.324-329.2003. DOI: https://doi.org/10.1128/JCM.41.1.324-329.2003
Wu, Q.; Hamilton, T.; Nelson, W. H.; Elliott, S.; Sperry, J. F.; Wu, M. Anal. Chem. 2001, 73, 3432–3440. DOI: https://doi.org/10.1021/ac001268b. DOI: https://doi.org/10.1021/ac001268b
Sengupta, A.; Mujacic, M.; Davis, E. J. Anal. Bioanal. Chem. 2006, 386, 1379–1386. DOI: https://doi.org/10.1007/s00216-006-0711-z. DOI: https://doi.org/10.1007/s00216-006-0711-z
Zainudin, N.; Mohd Hairul, A. R.; Yusoff, M. M.; Tan, L. L.; Chong, K. F. Anal. Methods. 2014, 6, 7935–7941. DOI: https://doi.org/10.1039/C4AY01836B. DOI: https://doi.org/10.1039/C4AY01836B
Athamneh, A. I. M.; Alajlouni, R. A.; Wallace, R. S.; Seleem, M. N.; Sengera, R. S. Antimicrob. Agents Chemother. 2014, 58, 1302–1314. DOI: https://doi.org/10.1128/aac.02098-13. DOI: https://doi.org/10.1128/AAC.02098-13
Guler, Z.; Sarac, A. S. eXPRESS Polym. Lett. 2016, 10, 96–110. DOI: https://doi.org/10.3144/expresspolymlett.2016.11
Lian, Y.; He, F.; Wang, H.; Tong, F. Biosens. Bioelectron. 2015, 65, 314–319. DOI: https://doi.org/10.1016/j.bios.2014.10.017
Upadhyayula, V. K. K.; Deng, S.; Mitchell, M. C.; Smith, G. B. Sci. Total Environ. 2009, 408, 1–13. DOI: https://doi.org/10.1016/j.scitotenv.2009.09.027. DOI: https://doi.org/10.1016/j.scitotenv.2009.09.027
Radke, S. M.; Alocilja, E. C. Biosens. Bioelectron. 2005, 20, 1662–1667. DOI: https://doi.org/10.1016/j.bios.2004.07.021. DOI: https://doi.org/10.1016/j.bios.2004.07.021
García-Aljaro, C.; Cella, L. N.; Shirale, D. J.; Park, M.; Muñoz, F. J.; Yates, M. V.; Mulchandani, A. Biosens. Bioelectron. 2010, 26, 1437–1441. DOI: https://doi.org/10.1016/j.bios.2010.07.077. DOI: https://doi.org/10.1016/j.bios.2010.07.077
Demattê, J. A. M.; Ramirez-Lopez, L.; Marques, K. P. P.; Rodella, A. A. Geoderma. 2017, 288, 8–22. DOI: https://doi.org/10.1016/j.geoderma.2016.11.013
Hayat, A.; Marty, J. L. Front. Chem. 2014. DOI: https://doi.org/10.3389/fchem.2014.00041. DOI: https://doi.org/10.3389/fchem.2014.00041
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Copyright (c) 2025 Elia Velázquez, Oscar Monroy Hermosillo , Florina Ramírez Vives , Michel Picquart , Israel Morales-Reyes , Paulina Hernández Garcés , Antonio Abad Sánchez , Eugenio Gómez Reyes Gómez Reyes , Nikola Batina
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