College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China; Laboratory of Quality & Safety Risk Assessment for Agro-products (Yangling), Ministry of Agriculture, Yangling, Shaanxi 712100, China; National Engineering Research Center of Agriculture Integration Test (Yangling), Yangling, Shaanxi 712100, China.
College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi 712100, China; Laboratory of Quality & Safety Risk Assessment for Agro-products (Yangling), Ministry of Agriculture, Yangling, Shaanxi 712100, China; National Engineering Research Center of Agriculture Integration Test (Yangling), Yangling, Shaanxi 712100, China.
Food Res Int. 2018 Dec;114:104-113. doi: 10.1016/j.foodres.2018.07.065. Epub 2018 Aug 2.
An approach based on multifunctional fluorescent magnetic nanoparticles was proposed for the enrichment and identification of Alicyclobacillus spp. in apple juice simultaneously. The prepared FeO magnetic particles (MNPs) were modified by the sol-gel process and a silica shell was formed to improve the reactivity, and then the obtained MNPs@SiO-SH nanoparticles were conjugated with Thioglycolic acid functionalized CdTe/CdS QDs via thiols chemistry. The characteristic evaluation results indicated that the MNPs-QD nanocomposites exhibited good magnetic properties and optical characterization. The polyclonal anti-Alicyclobacillus IgG antibody was immobilized onto the surface of MNPs-QD materials via esterification reactions. The maximum antibody immobilization capacity was 119.62 μg/mg and the adsorption reaction could be accomplished in 60 min. The adsorption process could be represented by Langmuir model and pseudo-second order kinetics equation, respectively. Based on the high immunocapture efficiency and sensitive fluorescence characteristics, the obtained MNPs-QDs-antibody conjugates could be applied to recognize the contamination of Alicyclobacillus spp. and a quantitative analysis method was established for target cells detection. The minimum quantitative limit was 10 CFU (colony forming unit)/mL and the testing process could be completed in 90 min. The results indicated that the MNPs-QDs-antibody conjugates can be successfully applied for immunocapture and detection of Alicyclobacillus spp. in apple juice. That is to say, the developed MNPs-QDs-antibody conjugates have exhibit more attractive and great potential for the immunocapture and recognition of target bacteria, fully demonstrated a new method for enrichment and rapid detection of Alicyclobacillus spp. in fruit juices.
提出了一种基于多功能荧光磁性纳米粒子的方法,用于同时富集和鉴定苹果汁中的环状芽孢杆菌。通过溶胶-凝胶法对制备的 FeO 磁性颗粒(MNPs)进行改性,形成硅壳以提高反应性,然后通过巯基化学将所得的 MNPs@SiO-SH 纳米粒子与巯基乙磺酸功能化的 CdTe/CdS QDs 偶联。特征评估结果表明,MNPs-QD 纳米复合材料具有良好的磁性和光学特性。多克隆抗环状芽孢杆菌 IgG 抗体通过酯化反应固定在 MNPs-QD 材料的表面上。最大抗体固定化容量为 119.62μg/mg,吸附反应可在 60min 内完成。吸附过程分别可以用 Langmuir 模型和拟二级动力学方程来表示。基于高免疫捕获效率和灵敏的荧光特性,所获得的 MNPs-QDs-抗体偶联物可用于识别环状芽孢杆菌的污染,并建立了用于目标细胞检测的定量分析方法。最低定量限为 10 CFU(集落形成单位)/mL,检测过程可在 90min 内完成。结果表明,MNPs-QDs-抗体偶联物可成功应用于苹果汁中环状芽孢杆菌的免疫捕获和检测。也就是说,开发的 MNPs-QDs-抗体偶联物在免疫捕获和识别目标细菌方面具有更具吸引力和巨大潜力,充分展示了一种用于果汁中环状芽孢杆菌的富集和快速检测的新方法。