A Bipolar Electrode-Electrochemiluminescence Sensor for Sensitive Detection of Escherichia coli O157:H7 Based on Loop-Mediated Isothermal Amplification

IF 3 3区农林科学 Q2 FOOD SCIENCE & TECHNOLOGY

Food Analytical Methods Pub Date : 2025-03-25 DOI:10.1007/s12161-025-02795-x

Yan Qi, Fengyang Wang, Yanjun Jiang, Lei Ji, Xiaohui Xiong, Yuanjian Liu

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引用次数: 0

Abstract

Escherichia coli O157:H7 (E. coli O157:H7) is a foodborne pathogen with high pathogenicity and lethality, capable of producing substantial amounts of Vero toxin (VT), posing a significant threat to human health. Consequently, a platform based on the bipolar electrode-electrochemiluminescence (BPE-ECL) and loop-mediated isothermal amplification (LAMP) was constructed for the detection of E. coli O157:H7. Firstly, LAMP technology is utilized to efficiently obtain the amplified product (double-stranded DNA, dsDNA) of E. coli O157:H7. On the one hand, SYBR Green was added to the amplified product to detect E. coli O157:H7 by colorimetric determination. The concentration of 10 CFU mL⁻¹ E. coli O157:H7 can only obtain qualitative analysis, to determine the presence of the target bacteria. On the other hand, methylene blue (MB) molecules were embedded in the dsDNA through electrostatic interactions. Then, by dropwise adding the mixture onto the cathode of the BPE, the ECL signal of Ru(bpy)₃²⁺ onto the anode was generated. Specifically, the BPE-ECL sensor displayed superior sensitivity for E. coli O157:H7 with a detection limit of 10 CFU mL⁻¹ and a wide linear concentration range of 10¹–10⁶ CFU mL⁻¹. In summary, this sensor can effectively enhance the sensitivity and reliability of detection when compared to traditional colorimetric methods, and achieve qualitative and quantitative detection of E. coli O157:H7 by measuring the anode ECL signal of Ru(bpy)₃²⁺.

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基于环介导等温扩增的灵敏检测大肠杆菌O157:H7的双极电极电化学发光传感器

大肠杆菌O157:H7 （E. coli O157:H7）是一种具有高致病性和致死率的食源性病原体，能够产生大量Vero毒素（VT），对人类健康构成重大威胁。为此，建立了双极电极电化学发光（BPE-ECL）和环介导等温扩增（LAMP）检测大肠杆菌O157:H7的检测平台。首先，利用LAMP技术高效获得大肠杆菌O157:H7的扩增产物（双链DNA， dsDNA）。一方面，在扩增产物中加入SYBR Green，用比色法检测大肠杆菌O157:H7；浓度为10 CFU mL−1的大肠杆菌O157:H7只能获得定性分析，以确定目标菌的存在。另一方面，亚甲基蓝（MB）分子通过静电相互作用嵌入dsDNA中。然后，通过将混合物滴加到BPE的阴极上，在阳极上产生Ru(bpy)32+的ECL信号。具体而言，BPE-ECL传感器对大肠杆菌O157:H7的检测限为10 CFU mL−1，线性浓度范围为101-106 CFU mL−1，具有较高的灵敏度。综上所述，与传统比色法相比，该传感器可有效提高检测的灵敏度和可靠性，通过测量Ru(bpy)32+的阳极ECL信号，实现对大肠杆菌O157:H7的定性和定量检测。

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来源期刊

Food Analytical Methods 农林科学-食品科技

CiteScore

6.00

自引率

3.40%

发文量

244

审稿时长

3.1 months

期刊介绍： Food Analytical Methods publishes original articles, review articles, and notes on novel and/or state-of-the-art analytical methods or issues to be solved, as well as significant improvements or interesting applications to existing methods. These include analytical technology and methodology for food microbial contaminants, food chemistry and toxicology, food quality, food authenticity and food traceability. The journal covers fundamental and specific aspects of the development, optimization, and practical implementation in routine laboratories, and validation of food analytical methods for the monitoring of food safety and quality.