| 基于dsDNA-CuNCs的荧光ELISA检测牛奶中的E.coli O157:H7 |
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| 引用本文： | 曹文凯,山 珊,刘道峰,彭 娟,邢克宇,赖卫华. 基于dsDNA-CuNCs的荧光ELISA检测牛奶中的E.coli O157:H7[J]. 食品与机械, 2023, 39(9): 38-43,94 |
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| 作者姓名： | 曹文凯 山 珊 刘道峰 彭 娟 邢克宇 赖卫华 |
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| 作者单位： | 南昌大学食品学院，江西 南昌 330047;江西师范大学生命科学学院，江西 南昌 330022;江西省疾病预防控制中心，江西 南昌 330029；江西省食源性疾病诊断溯源重点实验室，江西 南昌 330029;长沙理工大学食品科学与生物工程学院，湖南 长沙 410114 |
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| 基金项目： | 国家自然科学基金地区项目（编号：82260644）；江西省高层次高技能领军人才培养工程项目（2021年度） |
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| 摘 要： | 目的:建立了一种灵敏检测牛奶中大肠杆菌O157:H7的新型荧光酶联免疫吸附方法。方法:以碱性磷酸酶水解焦磷酸盐-Cu2+配位复合物，释放出Cu2+形成铜纳米簇作为信号报告探针，通过对关键因素Cu2+浓度、焦磷酸盐浓度、DNA模板的浓度和长度进行优化。结果:在最优条件下，大肠杆菌O157:H7的菌数为5×104～1×108 CFU/mL时，与荧光信号值有较好的线性关系(R2=0.980 8),最低检出限为2.4×104 CFU/mL。结论:该方法成功地应用于低脂牛奶和脱脂牛奶样本中大肠杆菌O157:H7的测定，平均回收率为92.2%～108.5%,相对标准偏差为4.9%～10.4%。
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| 关 键 词： | 酶联免疫吸附 铜纳米簇 大肠杆菌O157:H7 检测 |
A fluorescence ELISA based on dsDNA-CuNCs for the detection of E. coli O157:H7 in milk |
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| CAO Wenkai,SHAN Shan,LIU Daofeng,PENG Juan,XING Keyu,LAI Weihua. A fluorescence ELISA based on dsDNA-CuNCs for the detection of E. coli O157:H7 in milk[J]. Food and Machinery, 2023, 39(9): 38-43,94 |
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| Authors: | CAO Wenkai SHAN Shan LIU Daofeng PENG Juan XING Keyu LAI Weihua |
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| Affiliation: | College of Food Science, Nanchang University, Nanchang, Jiangxi 330047, China;College of Life Science, Jiangxi Normal University, Nanchang, Jiangxi 330022, China;Jiangxi Provincial Center for Disease Control and Prevention, Nanchang, Jiangxi 330029, China;Jiangxi Provincial Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Nanchang, Jiangxi 330029, China;School of Food Science and Bioengineering, Changsha University of Science & Technology, Changsha, Hunan 410114, China |
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| Abstract: | Objective: This study aimed to developed a novel fluorescent enzyme-linked immunosorbent assay method for the sensitive detection of E. coli O157:H7 in milk samples. Methods: The pyrophosphate-Cu2+ coordination complex was hydrolyzed by alkaline phosphatase, and Cu2+ was released from copper nanoclusters as signal reporting probes. Key factors such as Cu2+ concentration, pyrophosphate concentration, and DNA template concentration and length were optimized. Results: Under the control of the optimized conditions, the method exhibited a good linear relationship (R2=0.980 8) with the target E. coli O157:H7 in the range of 5×104~1×108 CFU/mL. The limit of detection was 2.4×104 CFU/mL. Conclusion: The method was successfully applied to the determination of E. coli O157:H7 in samples of low-fat milk and skim milk. The average recovery was 92.2% ~ 108.5% with relative standard deviation 4.9% ~ 10.4%. |
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| Keywords: | enzyme-linked immunosorbent assay copper nanocluster E. coli O157:H7 detection |
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