Zhao Yu, Ye Mingqiang, Chao Qiangguo, Jia Nengqin, Ge Yu, Shen Hebai
Department of Chemistry, Life and Environment Science College, Shanghai Normal University, People's Republic of China.
J Agric Food Chem. 2009 Jan 28;57(2):517-24. doi: 10.1021/jf802817y.
This paper reports a method that simultaneously detects three food-borne pathogenic bacteria, Salmonella typhimurium, Shigella flexneri, and Escherichia coli O157:H7, via an approach that combines magnetic microparticles for the enrichment and antibody-conjugated semiconductor quantum dots (QDs) as fluorescence markers. Using the water-in-oil reverse microemulsions method, the gamma-Fe(2)O(3) magnetic nanoparticles were coated with silica to empower the particles with high dispersibility and broad compatibility to biomacromolecules. The magnetic beads were then modified with amino silane, which could immobilize antibodies by glutaraldehyde treatment. The immunized magnetic beads and pathogenic bacteria formed "bead-cell" complexes in the enrichment procedure. QDs with different emission wavelengths (620, 560, and 520 nm) were immobilized with anti-S. typhimurium antibody, anti-S. flexneri antibody, and anti-E. coli O157:H7 antibody, respectively. Fluorescence microscope images and the fluorescence intensity of QDs labeled "sandwich" complexes (conjungated with antibodies against S. typhimurium, S. flexneri, and E. coli O157:H7, respectively) demonstrated that antibody-conjugated QDs could attach to the surface of bacterial cells selectively and specifically. In our method, we could detect food-borne pathogen bacteria in a food matrix at 10(-3) cfu/mL. We determined that a high concentration of proteins in food matrix would decrease the sensitivity of this method. This method, of which the detection procedures are completed within 2 h, can be applied to the rapid and cost-effective monitoring of bacterial contamination in food samples.
本文报道了一种通过结合用于富集的磁性微粒和作为荧光标记的抗体偶联半导体量子点(QDs),同时检测三种食源性病原体细菌——鼠伤寒沙门氏菌、宋内志贺氏菌和大肠杆菌O157:H7的方法。采用油包水反相微乳液法,用二氧化硅包覆γ-Fe(2)O(3)磁性纳米颗粒,使颗粒具有高分散性和对生物大分子的广泛兼容性。然后用氨基硅烷对磁珠进行修饰,氨基硅烷可通过戊二醛处理固定抗体。在富集过程中,免疫磁珠与病原菌形成“磁珠-细胞”复合物。分别用抗鼠伤寒沙门氏菌抗体、抗宋内志贺氏菌抗体和抗大肠杆菌O157:H7抗体固定发射波长不同(620、560和520 nm)的量子点。荧光显微镜图像以及量子点标记的“三明治”复合物(分别与抗鼠伤寒沙门氏菌、抗宋内志贺氏菌和抗大肠杆菌O157:H7抗体结合)的荧光强度表明,抗体偶联的量子点能够选择性且特异性地附着在细菌细胞表面。在我们的方法中,可以检测食品基质中浓度为10(-3) cfu/mL的食源性病原体细菌。我们确定食品基质中高浓度的蛋白质会降低该方法的灵敏度。该方法的检测过程在2小时内完成,可应用于食品样品中细菌污染的快速且经济高效的监测。