Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China.
Food Safety Key Laboratory of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China.
Biosens Bioelectron. 2018 Jun 1;107:266-271. doi: 10.1016/j.bios.2018.02.027. Epub 2018 Feb 15.
Here we innovate a portable and quantitative immunochromatographic assay (ICA) with a personal glucose meter (PGM) as readout for the detection of Escherichia coli O157:H7 (E. coli O157:H7). The carboxyl group coated FeO nanoparticles (MNPs) were synthesized by a one pot method and used as carriers of invertase and monoclonal antibody against E. coli O157:H7. Initially, the invertase and antibody double functionalized MNPs (Invertase-MNPs-IgG) conjugates were prepared and used as label probe in this assay system. Before laminating onto the baking card, the absorbent pad was soaked in sucrose solution and desiccated. MNPs produced brown band at the detection zone of the ICA when acting as direct labels. As they were also coupled with invertase, the invertase catalyzed the hydrolysis of sucrose on the absorbent pad into glucose, which was detected by the PGM. To increase the sensitivity, antibody functionalized MNPs were used to enrich E. coli O157:H7 from sample solution. The innovative aspect of this approach lies in the visualization and quantification of E. coli O157:H7 through Invertase-MNPs-IgG and the detection of glucose concentration using PGM. Although the feasibility is demonstrated using E. coli O157:H7 as a model analyte, this approach can be easily developed to be a universal analysis system and applied to detection of a wide variety of foodborne pathogens and protein biomarkers. This study proposed a qualitative and quantitative analysis device for the clinic diagnostics and food safety analysis.
我们创新了一种便携式定量免疫层析分析(ICA)方法,该方法使用个人血糖仪(PGM)作为读出器,用于检测大肠杆菌 O157:H7(E. coli O157:H7)。通过一锅法合成了带羧基的 FeO 纳米颗粒(MNPs),并将其用作蔗糖酶和针对大肠杆菌 O157:H7 的单克隆抗体的载体。最初,制备了蔗糖酶和抗体双功能化的 MNPs(Invertase-MNPs-IgG)缀合物,并将其用作该检测系统中的标记探针。在层压到烘烤卡之前,吸收垫被浸泡在蔗糖溶液中并干燥。MNPs 在作为直接标记物时在 ICA 的检测区产生棕色条带。由于它们还与蔗糖酶偶联,因此蔗糖酶将吸收垫上的蔗糖水解成葡萄糖,然后由 PGM 检测。为了提高灵敏度,使用抗体功能化的 MNPs 从样品溶液中富集大肠杆菌 O157:H7。该方法的创新之处在于通过 Invertase-MNPs-IgG 可视化和定量检测大肠杆菌 O157:H7,并使用 PGM 检测葡萄糖浓度。虽然使用大肠杆菌 O157:H7 作为模型分析物证明了其可行性,但该方法可以很容易地开发成为通用分析系统,并应用于检测各种食源性病原体和蛋白质生物标志物。本研究提出了一种用于临床诊断和食品安全分析的定性和定量分析装置。