Citation: LU Wen-Jie, ZHANG Xin-Ai, LI Chang-Feng, SHEN Jian-Zhong, JIANG Yu-Xiang, HUANG Chen-Yong. Electrochemical Immunosensor for Detection of E. coli in Urban Sludge Based on Dendrimer-encapsulated Au and Enhanced Gold Nanoparticle Labels[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(7): 1015-1021. doi: 10.11895/j.issn.0253-3820.160023 shu

Electrochemical Immunosensor for Detection of E. coli in Urban Sludge Based on Dendrimer-encapsulated Au and Enhanced Gold Nanoparticle Labels

1.
School of Energy and Power Engineering, Jiangsu University, Zhenjiang 212013, China;
2.
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China

Received Date: 10 January 2016
Revised Date: 11 April 2016

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21205051, 11072091), and the Key Project of Chinese Ministry of Education (No.201178)

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A novel electrochemical immunosensor was developed for detection of E. coli in urban sludge based on dendrimer-encapsulated Au and enhanced gold nanoparticle labels. p-Aminobenzoic acid (p-ABA) was electropolymerized on glassy carbon electrode (GCE) surface to introduce abundant carboxyl groups. G4-polyamidoamine dendrimers (PAMAM) were covalently attached onto electrode surface through the formation of amide bonds between amino groups of dendrimer and carboxyl groups of poly-p-ABA. Subsequently, Au(Ⅲ) ions were coordinated in the interior of dendrimer and then reduced to form gold nanoparticles (AuNPs). The resulting electrode (GCE/p-ABA/PAMAM(AuNPs)) provided numerous amino groups to allow highly dense immobilization of E. coli, and facilitated the improvement of electrochemical responses. The rabbit anti-E. coli polyclonal antibody (Ab₁) was captured by the electrode surface-confined E. coli, followed by the attachment of the enhanced gold nanoparticle labels (Ab₂-Au-Th) featuring thionine (Th) as signal-generating molecule. The analysis of E. coli was performed by electrochemical detection of Th in the bound labels on the electrode surface. Under the optimal experimental conditions, a linear relationship between the peak current of Th and the logarithmic value of E. coli concentration ranging from 1.0×10² cfu/mL to 1.0×10⁶ cfu/mL was obtained with a detection limit of 70 cfu/mL (S/N=3). The proposed strategy was also used to determine E. coli in samples obtained from waste water treatment plant, and the recoveries of standard additions were in the range of 89.4% to 105.8%. The results confirmed that the electrochemical immunoassay gave a useful protocol for E. coli analysis with high sensitivity, specificity and acceptable accuracy, and thus could potentially become a promising technique for estimating the feasibility of sludge recycling.
- Urban sludge,
- E. coli,
- Dendrimer,
- Functionalized gold nanoparticle labels,
- Electrochemical immunosensing
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沈阳化工大学材料科学与工程学院沈阳 110142