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一种基于适体传感器辅助表面增强拉曼光谱技术灵敏快速检测不同病原菌的通用方法。

A universal approach for sensitive and rapid detection of different pathogenic bacteria based on aptasensor-assisted SERS technique.

作者信息

Zhu Xiaoying, Ning Ying, Zhang Zeshuai, Wen Youqing, Zhao Yuwen, Wang Haixia

机构信息

College of Pharmaceutical Engineering of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, No. 10, Poyanghu Road, Jinghai District, Tianjin, 301617, China.

State Key Laboratory of Component-Based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.

出版信息

Anal Bioanal Chem. 2023 Mar;415(8):1529-1543. doi: 10.1007/s00216-023-04551-8. Epub 2023 Jan 27.

Abstract

An assembled-aptasensor based on FeO@Au@Ag nanocomposites grafting onto the gold foil was prepared, which can be developed into a universal approach for sensitive and rapid detection of various pathogenic bacteria, such as Escherichia coli (E. coli), Salmonella typhimurium (S. typhimurium), Staphylococcus aureus (S. aureus), Listeria monocytogenes (L. monocytogenes), Pseudomonas aeruginosa (P. aeruginosa), and Shigella flexneri (S. flexneri). Firstly, the gold foil paper was modified with thiolated capture probe and SERS tag in proportion, and at the same time, the specific thiolated aptamer probe for corresponding pathogenic bacteria was fixed with FeO@Au@Ag nanocomposites. An obvious Raman signal can be subsequently increased about 10 times by the external electromagnetic field enhancement at the "hot spots" caused by the hybridization of aptamer and capture probe. But in the presence of target pathogenic bacteria, Raman intensity will decrease as FeO@Au@Ag nanocomposites are dissociated from gold foil. Thus, all of the concentrations of the six kinds of pathogenic bacteria both in PBS and liquorice extract showed an obvious negative linear correlation with the Raman intensity of SERS tag in the range of 10-10 CFU/mL with detection limits were all lower than 10 CFU/mL. And there was no significant difference between our method and the plate counting method. Besides, the assembled-aptasensor had superior specific recognition ability even in the mixed interfering bacteria. Our study showed that this assembled-aptasensor had good specific detection ability to a variety of foodborne pathogens based on magnetic field-assisted SERS technique, which can be used for rapid and sensitive detection of a variety of pathogens in complex substrates.

摘要

制备了一种基于嫁接到金箔上的FeO@Au@Ag纳米复合材料的组装适体传感器,该传感器可发展成为一种灵敏、快速检测各种病原菌的通用方法,如大肠杆菌、鼠伤寒沙门氏菌、金黄色葡萄球菌、单核细胞增生李斯特菌、铜绿假单胞菌和宋内志贺氏菌。首先,将金箔纸按比例用硫醇化捕获探针和表面增强拉曼散射(SERS)标签进行修饰,同时,将针对相应病原菌的特异性硫醇化适体探针与FeO@Au@Ag纳米复合材料固定。随后,在适体与捕获探针杂交产生的“热点”处,外部电磁场增强可使明显的拉曼信号增加约10倍。但在目标病原菌存在的情况下,随着FeO@Au@Ag纳米复合材料从金箔上解离,拉曼强度会降低。因此,在10 - 10 CFU/mL范围内,六种病原菌在磷酸盐缓冲盐水(PBS)和甘草提取物中的浓度均与SERS标签的拉曼强度呈明显的负线性相关,检测限均低于10 CFU/mL。并且我们的方法与平板计数法之间没有显著差异。此外,即使在混合干扰细菌存在的情况下,组装适体传感器仍具有优异的特异性识别能力。我们的研究表明,这种基于磁场辅助SERS技术的组装适体传感器对多种食源性病原体具有良好的特异性检测能力,可用于复杂基质中多种病原体的快速灵敏检测。

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