Luo Yiheng, Dou Wenchao, Zhao Guangying
Food Safety Key Laboratory of Zhejiang Province, College of Food Science and Biotechnology, Zhejiang Gongshang University, 18 Xuezheng Street, Xiasha Higher Education Zone, Hangzhou, Zhejiang, 310018, China.
Anal Bioanal Chem. 2017 Jul;409(17):4139-4147. doi: 10.1007/s00216-017-0361-3. Epub 2017 Apr 21.
In this article, a facile and sensitive electrochemical method for quantification of Salmonella Pullorum and Salmonella Gallinarum (S. Pullorum and S. Gallinarum) was established by monitoring glucose consumption with a personal glucose meter (PGM). Antibody-functionalized magnetic nanoparticles (IgG-MNPs) were used to capture and enrich S. Pullorum and S. Gallinarum, and IgG-MNPs-S. Pullorum and IgG-MNPs-S. Gallinarum complexes were magnetically separated from a sample using a permanent magnet. The trace tag was prepared by loading polyclonal antibodies and high-content glucose oxidase on amino-functionalized silica nanoparticles (IgG-SiNPs-GOx). With a sandwich-type immunoassay format, IgG-SiNPs-GOx were added into the above mixture solution and conjugated to the complexes, forming sandwich composites IgG-MNPs/S. Pullorum and S. Gallinarum/IgG-SiNPs-GOx. The above sandwich composites were dispersed in glucose solution. Before and after the hydrolysis of glucose, the concentration of glucose was measured using PGM. Under optimal conditions, a linear relationship between the decrease of glucose concentration and the logarithm of S. Pullorum and S. Gallinarum concentration was obtained in the concentration range from 1.27 × 10 to 1.27 × 10 CFU mL, with a detection limit of 7.2 × 10 CFU mL (S/N = 3). This study provides a portable, low-cost, and quantitative analytical method for bacteria detection; thus, it has a great potential in the prevention of disease caused by S. Pullorum and S. Gallinarum in poultry. Graphical abstract A schematic illustration of the fabrication process of IgG-SiNPs-GOD nanomaterials (A) and IgG-MNPs (B) and experimental procedure of detection of S. Pullorum and S. Gallinarum using GOD-functionalized silica nanospheres as trace tags based on PGM (C).
在本文中，通过使用个人血糖仪（PGM）监测葡萄糖消耗，建立了一种简便灵敏的电化学方法来定量检测鸡白痢沙门氏菌和鸡伤寒沙门氏菌（鸡白痢沙门氏菌和鸡伤寒沙门氏菌）。抗体功能化磁性纳米颗粒（IgG-MNPs）用于捕获和富集鸡白痢沙门氏菌和鸡伤寒沙门氏菌，使用永久磁铁从样品中磁性分离IgG-MNPs-鸡白痢沙门氏菌和IgG-MNPs-鸡伤寒沙门氏菌复合物。通过将多克隆抗体和高含量葡萄糖氧化酶负载到氨基功能化二氧化硅纳米颗粒（IgG-SiNPs-GOx）上来制备痕量标记物。采用夹心型免疫分析形式，将IgG-SiNPs-GOx加入上述混合溶液中并与复合物结合，形成夹心复合物IgG-MNPs/鸡白痢沙门氏菌和鸡伤寒沙门氏菌/IgG-SiNPs-GOx。将上述夹心复合物分散在葡萄糖溶液中。在葡萄糖水解前后，使用PGM测量葡萄糖浓度。在最佳条件下，葡萄糖浓度的降低与鸡白痢沙门氏菌和鸡伤寒沙门氏菌浓度的对数在1.27×10至1.27×10 CFU mL的浓度范围内呈线性关系，检测限为7.2×10 CFU mL（S/N = 3）。本研究提供了一种用于细菌检测的便携式、低成本定量分析方法；因此，它在预防家禽由鸡白痢沙门氏菌和鸡伤寒沙门氏菌引起的疾病方面具有巨大潜力。图形摘要 IgG-SiNPs-GOD纳米材料（A）和IgG-MNPs（B）的制备过程示意图以及基于PGM使用GOD功能化二氧化硅纳米球作为痕量标记物检测鸡白痢沙门氏菌和鸡伤寒沙门氏菌的实验步骤（C）。
