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磁珠纳米探针结合免疫层析法联级信号放大快速检测奶制品中的两种致病菌
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Abstract:
阪崎氏肠杆菌和金黄色葡萄球菌是乳制品中致病率较高且常见的检测项目，本研究首先通过免疫磁分离技术实现目标菌的有效富集，然后通过新型的纳米荧光材料量子点及生物素–链霉亲和素体系结合免疫层析法实现阪崎氏肠杆菌和金黄色葡萄球菌联级信号放大和超灵敏联合检测。通过对研究整体的相关优化，两种菌检测灵敏度均可达到2 × 103 CFU/mL，相比传统的免疫层析法检测灵敏度提高了近2000倍。可实现同时超灵敏检测食源性致病菌的，为食品安全快速高灵敏准确检测大分子目标物提供了新的思路。
Enterobacter sakazakii and Staphylococcus aureus are common detection items with high pathogenicity in dairy products. In this study, we first achieved effective enrichment of target bacteria through immunomagnetic separation, realized signal amplification and sensitive detection of Enterobacter sakazakii and Staphylococcus aureus through quantum dots and biotin streptavidin system combined with immunochromatography. By optimizing the overall research, both bacterial detection sensitivities can reach 2 × 103 CFU/mL, which increases the detection sensitivity by nearly 2000 times compared to traditional immunochromatography. It can achieve simultaneous sensitive detection of foodborne pathogens, providing a new approach for food safety to quickly, sensitively, and accurately detect large molecular targets.
| [1] | 王翔, 等. 阪崎肠杆菌(克罗诺杆菌属)分子检测方法研究进展[J]. 食品科学, 2011, 32(13): 350-354. |
| [2] | 徐高杰, 等. 河南省9类食品中克罗诺杆菌属的污染检测[J]. 中国卫生检验杂志, 2022, 32(13): 1549-1551+1555. |
| [3] | Parra-Flores, J., et al. (2018) Cronobacter sakazakii and Microbiological Parameters in Dairy Formulas Associated with a Food Alert in Chile. Frontiers in Microbiology, 9, Article 1708. https://doi.org/10.3389/fmicb.2018.01708 |
| [4] | 覃昱. 应用免疫磁珠分离及LAMP技术快速检测配方奶粉中克罗诺杆菌[D]: [博士学位论文]. 广州: 南方医科大学, 2014. |
| [5] | 洪秀琴. 食品中金黄色葡萄球菌检验技术探讨[J]. 中国食品工业, 2022(17): 78-81. |
| [6] | 陈洪升. 金黄色葡萄球菌氨基糖苷类、喹诺酮耐药基因的研究[D]: [硕士学位论文]. 广州: 广东药学院, 2015. |
| [7] | 董彬, 封丽霞, 赵婷婷. 生乳中金黄色葡萄球菌的风险评估[J]. 食品安全导刊, 2018(28): 64-65. |
| [8] | Szczuka, E., et al. (2022) Occurrence and Characteristics of Staphylococcus aureus Isolated from Dairy Products. Molecules, 27, Article 4649. https://doi.org/10.3390/molecules27144649 |
| [9] | 柯振华. 餐饮食品中蜡样芽孢杆菌的实时荧光PCR检测方法研究[J]. 福建轻纺, 2020(4): 19-23. |
| [10] | 顾培科. 免疫磁分离结合胶体金免疫层析法快速检测大肠杆菌O157: H7[J]. 大家健康(学术版), 2015, 9(2): 155-156. |
| [11] | 林吉恒, 等. 免疫磁珠分离技术在食源性致病菌检测中的应用[J]. 食品安全质量检测学报, 2019, 10(18): 5998- 6005. |
| [12] | Cheng, X., et al. (2023) Simultaneous Detection of Foodborne Pathogenic Bacteria in Milk by Fluorescence Immunoassay. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 285, Article ID: 121830.
https://doi.org/10.1016/j.saa.2022.121830 |
| [13] | 肖有玉, 卢菲, 张弛, 王辰, 祝晨辰, 李东元, 唐伟. 一种新型定量检测恩诺沙星量子点免疫层析方法的建立[J]. 现代食品科技, 2020, 36(12): 252-258. |
| [14] | Xue, L., et al. (2018) An Ultrasensitive Fluorescent Biosensor Using High Gradient Magnetic Separation and Quantum Dots for Fast Detection of Foodborne Pathogenic Bacteria. Sensors and Actuators B: Chemical, 265, 318-325.
https://doi.org/10.1016/j.snb.2018.03.014 |
| [15] | Zhao, Y., et al. (2009) Simultaneous Detection of Multifood-Borne Pathogenic Bacteria Based on Functionalized Quantum Dots Coupled with Immunomagnetic Separation in Food Samples. Journal of Agricultural and Food Chemistry, 57, 517-524. https://doi.org/10.1021/jf802817y |
| [16] | 崔希, 等. 免疫磁分离结合胶体金免疫层析法快速检测大肠杆菌O157:H7 [J]. 分析化学, 2013, 41(12): 1812-1816. |
| [17] | 彭鹏. 硝基呋喃类兽药胶体金免疫层析检测方法研究[D]: [硕士学位论文]. 天津: 天津科技大学, 2016. |
| [18] | 宋翠翠, 等. 25-羟基维生素D化学发光检测试剂盒的研发[J]. 生物学杂志, 2017, 34(5): 108-112. |
| [19] | Zió？kowski, H., et al. (2014) Influence of Oral Co-Administration of a Preparation Containing Calcium and Magnesium and Food on Enrofloxacin Pharmacokinetics. Research in Veterinary Science, 97, 99-104.
https://doi.org/10.1016/j.rvsc.2014.05.005 |
| [20] | Wang, Y., et al. (2017) Development and Application of an Immunoaffinity Column Clean-Up for Enrofloxacin Determination in Food Samples. Food & Agricultural Immunology, 28, 248-259.
https://doi.org/10.1080/09540105.2016.1263984 |
