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-Cu^(2+)配位复合物,释放出Cu^(2+)形成铜纳米簇作为信号报告探针,通过对关键因素Cu^(2+)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5×10^(4)~1×10^(8)CFU/mL时,与荧光信号值有较好的线性关系(R^(2)=0.9808),最低检出限为2.4×10^(4)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%~10^(8).5%,相对标准偏差为4.9%~10.4%。
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-Cu^(2+)coordination complex was hydrolyzed by alkaline phosphatase,and Cu^(2+)was released from copper nanoclusters as signal reporting probes.Key factors such as Cu^(2+)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(R^(2)=0.9808)with the target E.coli O157:H7 in the range of 5×10^(4)~1×10^(8)CFU/mL.The limit of detection was 2.4×10^(4)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%~10^(8).5%with relative standard deviation 4.9%~10.4%.
作者
曹文凯
山珊
刘道峰
彭娟
邢克宇
赖卫华
CAO Wenkai;SHAN Shan;IU Daofeng;PENG Juan;XING Keyu;LAI Weihua(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)
出处
《食品与机械》
CSCD
北大核心
2023年第9期38-43,94,共7页
Food and Machinery
基金
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)。
