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Amperometric immunosensor for the detection of Escherichia coli O157:H7 in food specimens.
Anal Biochem. 2012 Feb 01; 421(1):227-33.AB

Abstract

A novel, label-free amperometric immunosensor has been developed for the rapid detection of heat-killed Escherichia coli O157:H7 (E. coli O157:H7). This immunosensor was prepared as follows. First, the long-chain, amine-terminated alkanethiol 11-amino-1-undecanethiol hydrochloride (AUT) was self-assembled onto a gold electrode surface to form an ordered, oriented, compact, and stable monolayer possessing -NH(2) functional groups that could immobilize massive gold nanoparticles (GNPs). Next, chitosan-multiwalled carbon nanotubes-SiO(2)/thionine (CHIT-MWNTs-SiO(2)@THI) nanocomposites and GNPs multilayer films were prepared via layer-by-layer (LBL) assembly. The surface area enhancement from the LBL assembly of the multilayer films improves the stability of the immobilized CHIT-MWNTs-SiO(2)@THI. More important, the sensitivity and stability of the immunosensor can be enhanced proportionally to the quantity of the THI mediator immobilized on the electrode surface. Finally, the E. coli O157:H7 antibody (anti-E. coli O157:H7) was covalently bound to the GNP monolayer and its bioactivity was measured by enzyme-linked immunosorbent assay (ELISA). Transmission electron microscopy (TEM) was employed to characterize the morphology of the MWNTs, CHIT-MWNTs, and CHIT-MWNTs-SiO(2)@THI. Under optimal conditions, the calibration curve for heat-killed E. coli O157:H7 has a working range of 4.12×10(2)-4.12×10(5) colony-forming units (CFU)/ml, and the total assay time was less than 45 min.

Authors+Show Affiliations

Department of Clinical Laboratory Science, College of Medical Laboratory, Third Military Medical University, Chongqing 400038, People's Republic of China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22119072

Citation

Li, Yan, et al. "Amperometric Immunosensor for the Detection of Escherichia Coli O157:H7 in Food Specimens." Analytical Biochemistry, vol. 421, no. 1, 2012, pp. 227-33.
Li Y, Cheng P, Gong J, et al. Amperometric immunosensor for the detection of Escherichia coli O157:H7 in food specimens. Anal Biochem. 2012;421(1):227-33.
Li, Y., Cheng, P., Gong, J., Fang, L., Deng, J., Liang, W., & Zheng, J. (2012). Amperometric immunosensor for the detection of Escherichia coli O157:H7 in food specimens. Analytical Biochemistry, 421(1), 227-33. https://doi.org/10.1016/j.ab.2011.10.049
Li Y, et al. Amperometric Immunosensor for the Detection of Escherichia Coli O157:H7 in Food Specimens. Anal Biochem. 2012 Feb 1;421(1):227-33. PubMed PMID: 22119072.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Amperometric immunosensor for the detection of Escherichia coli O157:H7 in food specimens. AU - Li,Yan, AU - Cheng,Ping, AU - Gong,Junhua, AU - Fang,Lichao, AU - Deng,Jun, AU - Liang,Wenbin, AU - Zheng,Junsong, Y1 - 2011/11/04/ PY - 2011/08/10/received PY - 2011/10/25/revised PY - 2011/10/27/accepted PY - 2011/11/29/entrez PY - 2011/11/29/pubmed PY - 2012/5/25/medline SP - 227 EP - 33 JF - Analytical biochemistry JO - Anal Biochem VL - 421 IS - 1 N2 - A novel, label-free amperometric immunosensor has been developed for the rapid detection of heat-killed Escherichia coli O157:H7 (E. coli O157:H7). This immunosensor was prepared as follows. First, the long-chain, amine-terminated alkanethiol 11-amino-1-undecanethiol hydrochloride (AUT) was self-assembled onto a gold electrode surface to form an ordered, oriented, compact, and stable monolayer possessing -NH(2) functional groups that could immobilize massive gold nanoparticles (GNPs). Next, chitosan-multiwalled carbon nanotubes-SiO(2)/thionine (CHIT-MWNTs-SiO(2)@THI) nanocomposites and GNPs multilayer films were prepared via layer-by-layer (LBL) assembly. The surface area enhancement from the LBL assembly of the multilayer films improves the stability of the immobilized CHIT-MWNTs-SiO(2)@THI. More important, the sensitivity and stability of the immunosensor can be enhanced proportionally to the quantity of the THI mediator immobilized on the electrode surface. Finally, the E. coli O157:H7 antibody (anti-E. coli O157:H7) was covalently bound to the GNP monolayer and its bioactivity was measured by enzyme-linked immunosorbent assay (ELISA). Transmission electron microscopy (TEM) was employed to characterize the morphology of the MWNTs, CHIT-MWNTs, and CHIT-MWNTs-SiO(2)@THI. Under optimal conditions, the calibration curve for heat-killed E. coli O157:H7 has a working range of 4.12×10(2)-4.12×10(5) colony-forming units (CFU)/ml, and the total assay time was less than 45 min. SN - 1096-0309 UR - https://www.unboundmedicine.com/medline/citation/22119072/Amperometric_immunosensor_for_the_detection_of_Escherichia_coli_O157:H7_in_food_specimens_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(11)00715-9 DB - PRIME DP - Unbound Medicine ER -