采用溶剂热法合成上转换纳米颗粒(UCNPs)-Na YF4∶Yb,Er,进行表面功能化修饰,将其与己烯雌酚(DES)单克隆抗体偶联,制备荧光探针,以牛血清白蛋白-己烯雌酚(BSA-DES)偶联物和羊抗鼠二抗分别喷涂硝酸纤维膜,形成试纸条检测线(T线)和质控线(C线),建立了上转换发光免疫层析试纸条快速定量检测DES的方法。实验结果表明,此试纸条定量检测DES线性范围为25-10000 ng/m L(y=0.43927x-0.57647,R^2=0.996),检出限为20.84 ng/m L,单个样品检测时间为15 min,批内和批间变异系数均小于10%,特异性识别能力强,在37℃存放下7天检测值RSD的平均值约为15%。加标回收实验显示平均回收率为90.1%-115.2%,相对标准偏差小于5%,与高效液相色谱法有较好的一致性。
The upconversion nanoparticles(UCNPs) Na YF4∶ Yb,Er were synthesized by the solvent thermal method.Fluorescence probe was prepared by coupling the anti-diethylstilbestrol(DES) monoclonal antibody with UCNPs.The DES-labeled bovine serum albumin(BSA) conjugates and goat anti-mouse antibodies were sprayed onto the nitrocellulose membrane as the test line and the control line,respectively.The resultant fluorescence probes were introduced into the immunochromatographic strip for rapid determination of DES.The UCNPs-based immunochromatographic strips for determination of DES exhibited a good linear range from25 ng/m L to 10000 ng/m L(y = 0.43927x-0.57647,R2= 0.996),with a limit of detection of20.84 ng/m L.The detection time of the proposed UCNPs based immunochromatographic strips for each sample was only 15 min.The RSDs of the intra and inter assay were less than 10%.The strip showed excellent specificity to structural and functional analogues,and the average RSD of detection value after storage at 37℃ for 7 days was about 15%.The recovery experiment showed that the average recoveries of DES at 5 spiked concentration levels were 90.1%-115.2%,and the RSDs were below 5%.These results were consistent with that of HPLC.