Rapid colorimetric detection of Salmonella typhimuriumusing a selective filtration technique combined with antibody-magnetic nanoparticle nanocomposites
Rapid colorimetric detection of Salmonella typhimuriumusing a selective filtration technique combined with antibody-magnetic nanoparticle nanocomposites
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DOI：
10.1007/s00216-013-7497-6
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发表时间：
2014-01-01
影响因子：
4.3
通讯作者：
Kim, Min-Gon
中科院分区：
文献类型：
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作者：
Shim, Won-Bo;Song, Jeong-Eon;Kim, Min-Gon
对危害人类健康的病原菌进行检测，需要一种快速、方便、可靠、灵敏的检测方法。本研究建立了一种单克隆抗体(MAb)-磁性纳米颗粒（MNP）纳米复合材料的选择性过滤方法，用于快速灵敏的鼠伤寒沙门菌比色检测。该方法包括两个关键步骤：利用MAb-MNP纳米复合材料对细菌进行免疫磁分离和对纳米复合材料结合的细菌进行过滤。研究人员测量了残留在膜上的纳米复合材料的颜色信号，这反映了测试样品中细菌的数量。对于不同浓度的病原体(2 × 10（4)-2 × 10(1)个细胞），获得了8 ~ 90%的免疫磁捕获效率。方法优化后，在纯培养液和人工接种蔬菜（100个细胞/g）中均可检测到2 × 10(1)个鼠伤寒沙门氏菌细胞。结果表明，该方法对鼠伤寒沙门氏菌具有高度特异性，与其他致病菌无交叉反应，可在45 min内完成检测，与最近报道的基于磁颗粒固定抗体的方法相比，检测时间更短或相似。本研究也证明了单抗- mnp纳米复合材料在实际比色分析中的直接应用，而无需将细菌与磁珠分离。
Detection of pathogenic bacteria that pose a great risk to human health requires a rapid, convenient, reliable, and sensitive detection method. In this study, we developed a selective filtration method using monoclonal antibody (MAb)-magnetic nanoparticle (MNP) nanocomposites for the rapid and sensitive colorimetric detection of Salmonella typhimurium. The method contains two key steps: the immunomagnetic separation of the bacteria using MAb-MNP nanocomposites and the filtration of the nanocomposite-bound bacteria. Color signals from the nanocomposites remaining on the membrane were measured, which reflected the amount of bacteria in test samples. Immunomagnetic capture efficiencies of 8 to 90 % for various concentrations of the pathogen (2 x 10(4)-2 x 10(1) cells) were obtained. After optimization of the method, 2 x 10(1) cells of S. typhimurium in pure culture solution was detectable as well as in artificially inoculated vegetables (100 cells/g). The method was confirmed to be highly specific to S. typhimurium without cross-reaction to other pathogenic bacteria and could be concluded within 45 min, yielding results in a shorter or similar time period as compared with recently reported antibody immobilized on magnetic-particle-based methods. This study also demonstrated direct application of MAb-MNP nanocomposites without a dissociation step of bacteria from magnetic beads in colorimetric assays in practice.