Assembly of adaptive engineering aptamers for Escherichia coli and their application in all-in-one rapid detection

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2025-04-01 DOI:10.1007/s00604-025-07138-5

Mengyue Liu, Shicai Xu, Giovanna Marrazza

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Abstract

Multivalent engineering aptamers (multi-Apts) adaptive and specific to whole Escherichia coli (E. coli: CMCC 44102) cells based on hybridization chain reaction (HCR) were constructed for the first time. The dissociation constant (K_d) value for these multi-Apts was 48 nM, demonstrating a higher affinity than that of monovalent aptamers (mono-Apts) (K_d = 102 nM). Furthermore, the reaction equilibrium of multi-Apts was achieved within 20 min, with a reaction rate twice that of mono-Apts. To validate the exceptional performance of these multi-Apts, they were employed as recognition elements in conjunction with gold nanoparticles (AuNPs) colorimetric assays for the all-in-one rapid detection of E. coli. This method exhibited a linear detection range from 1 × 10² to 1 × 10⁷ CFU mL⁻¹, achieving a limit of detection (LOD) as low as 19 CFU mL⁻¹. The recovery of this method in tap water and milk were 85.7% to 101% and 81.8% to 98.2%, respectively. The results indicated that this method not only provided a wide detection range but also exhibited high sensitivity and accuracy. Additionally, this study demonstrated that multi-Apts possessed greater application potential in the detection of macromolecular substances such as bacteria. In short, this work provided a novel approach for rapid and all-in-one detection of E. coli in food.

Graphical Abstract

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大肠杆菌自适应工程适体的组装及其在一体化快速检测中的应用

利用杂交链反应（HCR）技术，首次构建了适应大肠杆菌（e.c oli: CMCC 44102）全细胞的多价工程适体（multi-Apts）。这些多apts的解离常数（Kd）值为48 nM，比单价适配体（single - apts）（Kd = 102 nM）具有更高的亲和力。此外，多apts在20 min内达到反应平衡，反应速率是单apts的两倍。为了验证这些多apts的卓越性能，将它们作为识别元件与金纳米颗粒（AuNPs）比色法结合使用，用于全面快速检测大肠杆菌。该方法的线性检测范围为1 × 102到1 × 10⁷CFU mL毒血症，检测限低至19 CFU mL毒血症。该方法在自来水和牛奶中的回收率分别为85.7% ~ 101%和81.8% ~ 98.2%。结果表明，该方法不仅检测范围广，而且具有较高的灵敏度和准确性。此外，本研究表明，multi-Apts在细菌等大分子物质的检测中具有更大的应用潜力。本研究为食品中大肠杆菌的快速、一体化检测提供了一种新的方法。图形抽象

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.