Immune Therapy Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yuseong-gu, Daejeon 305-806, South Korea.
Biosens Bioelectron. 2013 May 15;43:432-9. doi: 10.1016/j.bios.2012.12.052. Epub 2013 Jan 4.
We demonstrated the new antibody/gold nanoparticle/magnetic nanoparticle nanocomposites (antibody/AuNP/MNPs) and their application in the detection of the foodborne pathogen, Staphylococcus aureus (S. aureus), in milk. The nanocomposites were synthesized by coating the MNPs with bovine serum albumin (BSA) then adsorbing the AuNPs and anti-S. aureus antibodies on their surface. Using the completed immunomagnetic nanostructures, S. aureus inoculated in the milk sample was captured and isolated from the medium using the permanent magnet. The nanoparticle-bound cells as well as the unbound cells in the supernatant were enumerated via surface plating to evaluate the target binding capacity of the nanocomposites. The capture efficiencies of the antibody/AuNP/MNPs were 96% and 78% for S. aureus in PBS and the milk sample respectively, which were significantly higher than those of the antibody-coupled MNPs without any AuNP. The captured cells were also applied to the selective filtration system to produce color signals that were used for the detection of the target pathogen. During the filtration, the cells bound to the antibody/AuNP/MNPs remained on the surface of the membrane filter while unbound nanoparticles passed through the uniform pores of the membrane. After the gold enhancement, the cells-particles complex resting on the membrane surface rendered a visible color, and the signal intensity became higher as the target cell concentration increased. The detection limits of this colorimetric sensor were 1.5×10(3) and 1.5×10(5)CFU for S. aureus in PBS and the milk sample respectively. This sensing mechanism also had the high specificity for S. aureus over the other pathogens such as Escherichia coli, Listeria monocytogenes, and Salmonella enterica. The assay required only 40min to obtain the results. With the use of the appropriate antibodies, our immunomagnetic nanocomposites-based detection strategy can provide an easy, convenient, and rapid sensing method for a wide range of pathogens.
我们展示了新的抗体/金纳米粒子/磁性纳米粒子纳米复合材料(抗体/AuNP/MNPs)及其在检测食源性病原体金黄色葡萄球菌(S. aureus)中的应用。该纳米复合材料是通过用牛血清白蛋白(BSA)包覆 MNPs 然后在其表面吸附 AuNPs 和抗 S. aureus 抗体合成的。使用完成的免疫磁性纳米结构,从牛奶样品中接种的 S. aureus 可以使用永磁体从培养基中捕获和分离。通过表面平板计数来评估纳米复合材料的目标结合能力,对结合在纳米颗粒上的细胞以及上清液中未结合的细胞进行计数。抗体/AuNP/MNPs 在 PBS 和牛奶样品中对 S. aureus 的捕获效率分别为 96%和 78%,明显高于没有任何 AuNP 的抗体偶联 MNPs。捕获的细胞也被应用于选择性过滤系统以产生用于检测目标病原体的颜色信号。在过滤过程中,与抗体/AuNP/MNPs 结合的细胞保留在膜过滤器的表面上,而未结合的纳米颗粒则通过膜的均匀孔穿过。金增强后,停留在膜表面上的细胞-颗粒复合物呈现出可见的颜色,并且随着目标细胞浓度的增加,信号强度变得更高。这种比色传感器的检测限对于 PBS 中的 S. aureus 和牛奶样品中的 S. aureus 分别为 1.5×10(3)和 1.5×10(5)CFU。该传感机制对于金黄色葡萄球菌也具有比其他病原体(如大肠杆菌、单核细胞增生李斯特菌和肠炎沙门氏菌)更高的特异性。该测定仅需 40min 即可获得结果。通过使用适当的抗体,我们基于免疫磁性纳米复合材料的检测策略可以为广泛的病原体提供一种简单、方便、快速的传感方法。
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