Nanozyme-mediated multi-mode cascaded visual lateral flow assay platform of enrofloxacin based on truncated aptamer and one-pot isothermal amplification

IF 3.7 1区化学 Q1 CHEMISTRY, ANALYTICAL

Sensors and Actuators B: Chemical Pub Date : 2025-10-02 DOI:10.1016/j.snb.2025.138904

Peifang Chen, Mingwei Qin, Yu Li, Xiaodong Lu, Shuo Qi, Junsong Yang, Zhouping Wang

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Abstract

The widespread use of enrofloxacin (ENR) in livestock farming raises serious concerns for human health due to its persistent residues in food. Thus, this study innovatively developed a cascade-amplified "turn-on" lateral flow assay (LFA) for the highly sensitive and visual detection of ENR. The high-affinity truncated aptamer (E22), initially obtained through molecular docking simulation, as the recognition element. By combining one-pot rolling circle amplification (RCA) and restriction endonuclease, the generated single-stranded DNA were captured by functionalized gold-platinum nano-urchin probes (APNCs-PolyA-DP). Based on the generated initial colorimetric (CM) signal, the CM-LFA enabled quantitative detection of ENR in the range of 0.1-1000 ng/mL, with a visual detection limit (vLOD) of 0.1 ng/mL. Then, the peroxidase-like activity of APNCs catalyzed the TMB chromogenic reaction to produce enhanced colorimetric (EC) signals. Furthermore, the synergistic near-infrared photothermal (PT) properties of the oxidation product (oxTMB) and APNCs were utilized to construct a specific temperature mode. By leveraging the dual properties of oxTMB, the EC-LFA and PT-LFA achieved a wider linear range (0.01-1000 ng/mL) and an ultralow vLOD of 0.01 ng/mL, showing a 10-fold improvement over the CM-LFA. The application of the LFA to milk samples yielded overall recovery rates, ranging from 93.9% to 105.1%. Thus, this approach provides robust technical support for point-of-care testing (POCT) platforms and shows broad application prospects in food safety monitoring.

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基于截断适体和一锅等温扩增的纳米酶介导的恩诺沙星多模式级联视觉侧流检测平台

恩诺沙星（ENR）在畜牧业中的广泛使用引起了对人类健康的严重关注，因为它在食品中的持续残留。因此，本研究创新性地开发了一种级联放大“开启”横向流动试验（LFA），用于高灵敏度和视觉检测ENR。通过分子对接模拟初步获得的高亲和力截断适配体（E22）作为识别元件。结合单锅滚环扩增（RCA）和限制性内切酶，用功能化金-铂纳米海胆探针（APNCs-PolyA-DP）捕获生成的单链DNA。基于生成的初始比色（CM）信号，CM- lfa可在0.1 ~ 1000 ng/mL范围内定量检测ENR，视觉检出限（vLOD）为0.1 ng/mL。然后，apnc的过氧化物酶样活性催化TMB显色反应，产生增强的比色（EC）信号。此外，利用氧化产物（oxTMB）和apnc的协同近红外光热（PT）特性构建了特定的温度模式。通过利用oxTMB的双重特性，EC-LFA和PT-LFA实现了更宽的线性范围（0.01-1000 ng/mL）和超低vLOD (0.01 ng/mL)，比CM-LFA提高了10倍。LFA对牛奶样品的回收率为93.9% ~ 105.1%。因此，该方法为即时检测（POCT）平台提供了强有力的技术支持，在食品安全监测中具有广阔的应用前景。

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来源期刊

Sensors and Actuators B: Chemical 工程技术-电化学

CiteScore

14.60

自引率

11.90%

发文量

1776

审稿时长

3.2 months

期刊介绍： Sensors & Actuators, B: Chemical is an international journal focused on the research and development of chemical transducers. It covers chemical sensors and biosensors, chemical actuators, and analytical microsystems. The journal is interdisciplinary, aiming to publish original works showcasing substantial advancements beyond the current state of the art in these fields, with practical applicability to solving meaningful analytical problems. Review articles are accepted by invitation from an Editor of the journal.