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用于高通量检测食源性病原体的全细胞分子印迹荧光光子微球微阵列

Whole-Cell Molecularly Imprinted Fluorescent Photonic Microsphere Microarray for High-Throughput Detection of Foodborne Pathogens.

作者信息

Zhang Jingshuang, Liu Xiaomeng, Li Ziqiang, Li Xiang, Li Qianjin, Li Weiwei, Li Jianlin

机构信息

School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing 210023, China.

出版信息

Anal Chem. 2025 May 13;97(18):9779-9788. doi: 10.1021/acs.analchem.4c06821. Epub 2025 Apr 30.

Abstract

Rapid, sensitive, high-throughput, and cost-effective detection of multiplex foodborne pathogens is still challenging in public health. We designed a whole-cell imprinted microarray platform based on the surface of three-dimensional photonic microspheres for multiplex foodborne pathogenic bacteria using 3-formylphenylboric acid-functionalized silane as the functional monomer and fluorescein isothiocyanate-functionalized silane as the fluorescent monomer. After incubation with the multiplex pathogens, the fluorescent molecularly imprinted photonic microspheres can specifically capture the target pathogenic bacteria, and their fluorescence intensities can report the concentrations of the pathogens in samples. The new microsphere microarray showed wide linear detection ranges (10 to 10 CFU/mL for 10 to 10 CFU/mL for and 10 to 10 CFU/mL for O157:H7) and low limits of detection (LODs) (3 CFU/mL for 20 CFU/mL for and 1 CFU/mL for O157:H7) for multiplex pathogens. The new system does not require molecular probes such as antibody, aptamer, or DNA sequence and without enrichment culture and DNA amplification processes. The newly developed method has great potential applications in rapid, cost-effective, and high-throughput simultaneous detection of multiplex foodborne pathogens.

摘要

在公共卫生领域,快速、灵敏、高通量且经济高效地检测多种食源性病原体仍然具有挑战性。我们设计了一种基于三维光子微球表面的全细胞印迹微阵列平台,用于检测多种食源性病原体,该平台使用3-甲酰基苯硼酸功能化硅烷作为功能单体,异硫氰酸荧光素功能化硅烷作为荧光单体。与多种病原体孵育后,荧光分子印迹光子微球可特异性捕获目标病原菌,其荧光强度可反映样品中病原体的浓度。新型微球微阵列对多种病原体显示出宽线性检测范围(鼠伤寒沙门氏菌为10⁴至10⁹CFU/mL,肠炎沙门氏菌为10³至10⁸CFU/mL,O157:H7为10²至10⁷CFU/mL)和低检测限(鼠伤寒沙门氏菌为3CFU/mL,肠炎沙门氏菌为20CFU/mL,O157:H7为1CFU/mL)。该新系统不需要抗体、适体或DNA序列等高通量分子探针,也无需富集培养和DNA扩增过程。新开发的方法在快速、经济高效且高通量同时检测多种食源性病原体方面具有巨大的潜在应用价值。

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