基于LED光催化作用的可视化生物传感新方法快速超灵敏检测microRNA-21  

Rapid and ultrasensitive detection of microRNA-21 (miRNA-21) using visual analysis method based on LED photocatalysis

作　　者：于天晓 许红[1] 韩彦洁[1] 张瑶[1] 李美芳[1] 张敬蕊 腰利云 Yu Tianxiao;Xu Hong;Han Yanjie;Zhang Yao;Li Meifang;Zhang Jingrui;Yao Liyun(The Fourth Hospital of Shijiazhuang, Shijiazhuang 050000, China)

机构地区：[1]石家庄市第四医院

出　　处：《分析仪器》2019年第4期84-87,共4页Analytical Instrumentation

基　　金：河北省重点研发计划自筹项目(172777192)

摘　　要：目的:建立基于三交联(three-way junction,TWJ)DNA纳米结构及LED光催化作用的可视化生物传感方法快速超灵敏检测microRNA-21(miRNA-21)。方法:将生物素标记的miRNA-21捕获探针固定于亲和素化的96孔板上,捕获探针能够有效的结合靶物质miRNA-21,随后加入DNA检测探针P1、P2、P3,使其自组装形成TWJ DNA纳米结构。加入SYBR Green I后,dsDNA-SYBR Green I复合物在LED光照射下可催化3,3′,5,5′-四甲基联苯胺(TMB)产生显色反应,其吸光度与miRNA-21浓度成明显比例关系,从而可对miRNA-21浓度进行定量检测。结果:在最佳反应条件下,该方法最低检测限可达5.49pmol/L,充分说明该方法具有良好的灵敏度。考察方法的精密度,相对标准偏差RSD均在7.3%以下。该方法的回收率在79.8~128.8%之间,说明该方法具有良好的精密度和回收率。结论:本研究建立了一种基于TWJ DNA纳米结构及LED光催化作用的可视化生物传感分析方法,能够快速、超灵敏检测miRNA-21。该方法为miRNA的检测提供了新思路。This method provides a new idea for the detection of miRNA. Biotin labelled miRNA-21 capture probe was immobilized on a streptavidin labelled 96-well plate. The capture probe could effectively bind to target miRNA-21, then the detection probes P1, P2 and P3 were added to form a TWJ DNA nanostructures. Added with SYBR Green I, the dsDNA-SYBR Green I complex can catalyze 3,3′, 5,5′-tetramethylbenzidine(TMB) under LED irradiation, leading to color change. The absorbance was proportional to the concentration of miRNA21, and then the concentration of miRNA21 could be quantified. Under the optimized conditions, the lowest detection limit of the method was 5.49 pmol/L,indicating the method had perfect sensitivity.The relative standard deviations(RSD) were all less than 7.3%, the recoveries were 79.8%-128.8%, illustrating the method had good precision and recovery.

关 键 词：MIRNA-21 光催化作用 TWJ DNA纳米结构 

分 类 号：O643.32[理学—物理化学] O644.1[理学—化学] R440[医药卫生—诊断学]