Physiological Sensing Facility, Bindley Biosciences Center, Agricultural and Biological Engineering, Purdue University, 225 S. University Street, West Lafayette, IN 47907, USA.
Anal Bioanal Chem. 2011 Jan;399(3):1271-8. doi: 10.1007/s00216-010-4453-6. Epub 2010 Dec 7.
A rapid and sensitive method was developed here for separation and detection of multiple pathogens in food matrix by magnetic surface-enhanced Raman scattering (SERS) nanoprobes. Silica-coated magnetic probes (MNPs@SiO(2)) of ~100 nm in diameter were first prepared via the reverse microemulsion method using cetyltrimethylammonium bromide as a surfactant and tetraethyl orthosilicate as the silica precursor. The as-prepared MNPs@SiO(2) were functionalized with specific pathogen antibodies to first capture threat agents directly from a food matrix followed by detection using an optical approach enabled by SERS. In this scheme, pathogens were first immuno-magnetically captured with MNPs@SiO(2), and pathogen-specific SERS probes (gold nanoparticles integrated with a Raman reporter) were functionalized with corresponding antibodies to allow the formation of a sandwich assay to complete the sensor module for the detection of multiple pathogens in selected food matrices, just changing the kinds of Raman reporters on SERS probes. Here, up to two key pathogens, Salmonella enterica serovar Typhimurium and Staphylococcus aureus, were selected as a model to illustrate the probability of this scheme for multiple pathogens detection. The lowest cell concentration detected in spinach solution was 10(3) CFU/mL. A blind test conducted in peanut butter validated the limit of detection as 10(3) CFU/mL with high specificity, demonstrating the potential of this approach in complex matrices.
本研究开发了一种快速、灵敏的方法,通过磁表面增强拉曼散射(SERS)纳米探针分离和检测食品基质中的多种病原体。首先通过反相微乳液法制备直径约为 100nm 的二氧化硅包覆的磁性探针(MNPs@SiO2),该方法以十六烷基三甲基溴化铵(CTAB)为表面活性剂,正硅酸乙酯(TEOS)为硅源前驱体。所制备的 MNPs@SiO2 用特异性病原体抗体进行功能化,以直接从食品基质中捕获威胁剂,然后通过 SERS 实现的光学方法进行检测。在该方案中,病原体首先与 MNPs@SiO2 进行免疫磁捕获,然后将与病原体特异性的 SERS 探针(金纳米粒子与拉曼报告分子偶联)用相应的抗体进行功能化,以形成三明治检测,从而完成用于检测选定食品基质中多种病原体的传感器模块,只需改变 SERS 探针上拉曼报告分子的种类即可。在此,选择沙门氏菌 Typhimurium 和金黄色葡萄球菌两种关键病原体作为模型,说明了该方案用于多种病原体检测的可能性。在菠菜溶液中检测到的最低细胞浓度为 103 CFU/mL。在花生酱中进行的盲测验证了该方法的检测限为 103 CFU/mL,具有很高的特异性,表明该方法在复杂基质中具有应用潜力。