光子学报, 2019, 48 (7): 0717003, 网络出版: 2019-07-31
基于三明治SERS结构和酶剪切技术的肿瘤标志物miRNA-21的高灵敏检测
Highly Sensitive Detection of Tumor Marker miRNA-21 Based on Sandwich SERS Structure and Enzymatic Cleavage Technique
聚苯乙烯小球 表面增强拉曼散射 三明治结构 双链特异性剪切酶 肿瘤标志物 Polystyrene globules Surface-Enhanced Raman Scattering (SERS) Sandwich structure Double-stranded specific splicing enzyme Tumor markers
摘要
采用Langmiur-Bloggt膜技术和磁控溅射技术制备Ag覆盖聚苯乙烯小球六方密堆积阵列(Ag/PS HCA)基底, 再将合成的海胆状Au纳米粒子与4-巯基苯甲酸(4-Mercaptobenzoic acid, 4MBA)链接得到Au@4MBA探针, 然后将单链寡核苷酸DNA21分别与基底和Au@4MBA探针链接, 构建Au@4MBA-DNA21-Ag/PS HCA三明治结构, 在DNA21与miRNA-21杂交后使用双链特异性剪切酶(DSN)剪切DNA磷酸二酯键, 最后进行表面增强拉曼散射信号检测.实验结果表明, 基于上述三明治表面增强拉曼散射结构和酶剪切技术进行肿瘤标志物miRNA-21的检测, 在100 pmol·L-1到1 fmol·L-1的浓度范围内, 检测极限达到0.853 fmol·L-1, 具有极高的灵敏度和优良的特异性.
Abstract
An Ag-coated polystyrene pellet hexagonal close packed array (Ag/PS HCA) substrate was prepared by L-B film method and magnetron sputtering technique. And Au@4MBA probes were obtained by linking the synthesized urchin-like Au nanoparticles with 4-mercaptobenzoic acid (4MBA). Then the single-stranded oligonucleotide DNA21 was linked with the substrate and the Au@4MBA probes, respectively, to construct the Au@4MBA-DNA21-Ag/PS HCA sandwich structure. After DNA21 and miRNA-21 hybridization, the DNA phosphodiester bond is cleaved using a double-strand specific cleavage enzyme (DSN), and finally SERS signal detection was performed. The experimental results show that the detection scheme of the tumor marker miRNA-21 based on the above-mentioned sandwich SERS structure and enzymatic cleavage technology has extremely high sensitivity and excellent specificity, and the detection limit reaches 0.853 fmol·L-1. In addition, compared with real-time fluorescence quantitative polynucleotide chain reaction (RT-qPCR), our scheme not only has the same results also has higher sensitivity.
洑颢, 吕炜烽, 高永峰, 王哲, 邹祖全, 张鑫, 周骏. 基于三明治SERS结构和酶剪切技术的肿瘤标志物miRNA-21的高灵敏检测[J]. 光子学报, 2019, 48(7): 0717003. FU Hao, L Wei-feng, GAO Yong-feng, WANG Zhe, ZOU Zu-quan, ZHANG Xin, ZHOU Jun. Highly Sensitive Detection of Tumor Marker miRNA-21 Based on Sandwich SERS Structure and Enzymatic Cleavage Technique[J]. ACTA PHOTONICA SINICA, 2019, 48(7): 0717003.