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基于光可裂解水凝胶的用于检测副溶血性弧菌的磁弛豫开关生物传感器。

Magnetic relaxation switch biosensor for detection of Vibrio parahaemolyticus based on photocleavable hydrogel.

作者信息

Chen Le, Zhang Dongyu, Yang Fan, Shi Xizhi, Jiang Xiaohua, Hao Tingting, Zhang Qingqing, Hu Yufang, Wang Sui, Guo Zhiyong

机构信息

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, 315211, PR China.

State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, School of Marine Sciences, Ningbo University, Ningbo, 315211, PR China.

出版信息

Anal Chim Acta. 2025 Jan 22;1336:343516. doi: 10.1016/j.aca.2024.343516. Epub 2024 Dec 2.

Abstract

BACKGROUND

Foodborne pathogens, particularly Vibrio parahaemolyticus (VP) found in seafood, pose significant health risks, including abdominal pain, nausea, and even death. Rapid, accurate, and sensitive detection of these pathogens is crucial for food safety and public health. However, existing detection methods often require complex sample pretreatment, which limits their practical application. This study aims to overcome these limitations by developing a label-free magnetic relaxation switch (MRS) biosensor for the detection of VP, utilizing a photocleavable sol-gel phase transition system for improved efficiency and accuracy.

RESULTS

In this work, a tag-free magnetic relaxation switch (MRS) biosensor was designed for the detection of Vibrio parahaemolyticus (VP), based on a photocleavable sol-gel phase transition system. A large amount of lithium acyl hypophosphite (LAP), gold nanoparticles (AuNPs), and single-stranded DNA (ssDNA) loaded on the surface of TiCT MXene acted as the signal unit LAP-MXene@AuNPs-ssDNA. The pipette tip served as a reaction vessel, and when VP was present, Apt specifically captured VP and released the signal units. The released signal units were then injected into the low-field nuclear magnetic resonance (LF-NMR) test solution, a gel formed by crosslinking of disulfide bonds. The gel was cleaved by LAPs on the signal units under ultraviolet (UV) irradiation, triggering a gel-sol phase transition, which increased transverse relaxation time (T), thus enabling the detection of VP. Under the optimal experimental conditions, the linear range and detection limit for VP were 10 ∼ 10 CFU/mL and 10 CFU/mL, respectively.

SIGNIFICANCE AND NOVELTY

The simplified biometric identification process in the pipette tip reduces errors from multiple sample transfers, enhancing efficiency. The use of photocleavable hydrogel for signal output eliminates issues associated with magnetic material aggregation, significantly improving detection precision. The assay is of good selectivity, stability reproducibility, and convenience, having a broad application prospect in the rapid detection of pathogenic bacteria in the field.

摘要

背景

食源性病原体,尤其是海鲜中发现的副溶血性弧菌(VP),会带来重大健康风险,包括腹痛、恶心甚至死亡。快速、准确且灵敏地检测这些病原体对于食品安全和公共卫生至关重要。然而,现有的检测方法通常需要复杂的样品预处理,这限制了它们的实际应用。本研究旨在通过开发一种用于检测VP的无标记磁弛豫开关(MRS)生物传感器来克服这些限制,利用光可裂解溶胶-凝胶相变系统提高效率和准确性。

结果

在这项工作中,基于光可裂解溶胶-凝胶相变系统设计了一种用于检测副溶血性弧菌(VP)的无标记磁弛豫开关(MRS)生物传感器。负载在TiCT MXene表面的大量次磷酸锂酰(LAP)、金纳米颗粒(AuNPs)和单链DNA(ssDNA)充当信号单元LAP-MXene@AuNPs-ssDNA。移液器吸头用作反应容器,当存在VP时,适体特异性捕获VP并释放信号单元。然后将释放的信号单元注入低场核磁共振(LF-NMR)测试溶液中,该溶液是由二硫键交联形成的凝胶。在紫外线(UV)照射下,信号单元上的LAPs裂解凝胶,引发凝胶-溶胶相变,增加横向弛豫时间(T),从而实现对VP的检测。在最佳实验条件下,VP的线性范围和检测限分别为10~10⁶ CFU/mL和10³ CFU/mL。

意义与创新

移液器吸头中简化的生物识别过程减少了多次样品转移带来的误差,提高了效率。使用光可裂解水凝胶进行信号输出消除了与磁性材料聚集相关的问题,显著提高了检测精度。该检测方法具有良好的选择性、稳定性、重现性和便利性,在现场病原菌快速检测方面具有广阔的应用前景。

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