摘要
研究建立了基于量子点(quantum dots,QDs)的间接荧光免疫吸附测定技术(competitive fluorescence linked immunosorbentassay,cFLISA),并应用于牛奶中的氧氟沙星(ofloxacin,0FL)抗生素残留检测中。首先将量子点同抗氧氟沙星抗体通过生物素-亲和素系统进行连接,优化连接条件进行并测定连接效率。其次对cFLISA测定条件进行了优化,并确定了合适的检测时间≤60min。所建立方法的最低检测限(IC_(90)值)和IC_(50)值分别为0.25 ng·mL^(-1)和1.16 ng·mL^(-1)。cFLISA针对实际牛奶样品中氧氟沙星抗生素残留回收率测定结果为88.2%～105.3%,CV值为7.3%～10.9%,同传统酶联免疫吸附测定(ELLSA)方法相比十分接近。结果表明,本研究所建立的cFLISA方法可以替代传统的ELISA方法应用于牛奶中氧氟沙星抗生素残留的实际测定中。更重要的是,新方法对于针对不同药物残留的多重测定技术的建立具有一定的参考价值。
Using quantum dots（QDs） labels,a one -step competitive fluorescence linked immunosorbent assay（cFLISA） was developed for ofloxacin（OFL） detection as a model in this work.The QDs and OFL antibody was conjugated followed by characterization and optimization of the ligation accuracy,conjugation efficiency and assay conditions.The appropriate assay time with sufficient fluorescence intensity was less than 60 min.The limit of detennination（LOD,IC_（90）） and 50%inhibition value（IC_（50）） were 0.25 ng·mL^（-1） and 1.16 ng·mL^（-1）,respectively.Compared to competitive enzyme-linked immunosorbent assay（cELISA）,the satisfactory recoveries of the cFLISA from different spiked milk samples were from 88.2%to 105.3%with coefficients of variation（CV） of 7.3%～10.9%.The experimental results demonstrate that,as an alternative approach,the newly developed one-step cFLISA is capable of screening target drug in less time with identically desirable detection performance in comparison with cELISA.More importantly,the proposed detection mode would be valuable reference for further development of ideal multi-analysis system.
出处
《上海交通大学学报（农业科学版）》
2010年第6期541-545,共5页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
国家高技术研究发展计划(863)重点项目(2006AA10Z438
2007AA10Z436
2008AA100804)
上海市重点学科建设项目(B205)
