光子学报, 2019, 48 (7): 0717002, 网络出版: 2019-07-31   

基于SiC@Ag基底和银-生物素-链霉亲和素纳米聚集体双重SERS放大的miRNA-106a检测

Detection of miRNA-106a Based on Dual SERS Amplifications of SiC@Ag Substrate and Silver-biotin-streptavidin Nanoaggregates
作者单位
1 宁波大学 物理科学与技术学院 微电子科学与工程系, 浙江 宁波315211
2 宁波大学 医学院 浙江省病理生理学重点实验室, 浙江 宁波315211
摘要
基于SiC@Ag基底与Ag纳米颗粒的表面增强拉曼散射效应, 提出了利用银-生物素-链霉亲和素纳米聚集体二次表面增强拉曼散射放大的超灵敏miRNA-106a检测方案.首先, 将地高辛修饰的捕获DNA与固定在SiC@Ag基底上的抗地高辛链接, 制备SiC@Ag@anti-digoxin/digoxin-DNA基底; 将4-巯基苯甲酸(4MBA)标记的银纳米颗粒与修饰有氨基和生物素的探针DNA链接, 制备Ag@4MBA@DNA-biotin探针.然后将制备的基底、探针与待测miRNA-106a组成“三明治”结构, 获得表面增强拉曼散射信号放大.最后, 依次加入链霉亲和素和制备的探针, 形成银-生物素-链霉亲和素纳米聚集体, 实现检测信号的二次放大.实验结果表明, 利用SiC@Ag基底和银-生物素-链霉亲和素纳米聚集体双重表面增强拉曼散射放大, 可以实现miRNA-106a的超灵敏检测, 检测极限达到0.579 fmol/L, 对于肿瘤的早期诊断具有应用潜力.
Abstract
Based on the surface-enhanced Raman scattering enhancement effect of SiC@Ag substrate and Ag nanoparticles, an ultrasensitive miRNA-106a detection protocol was proposed by using the second surface-enhanced Raman scattering amplification of the silver-biotin-streptavidin nanoaggregates. First, SiC@Ag@anti-digoxin/digoxin-DNA substrate was prepared by specifically binding of a capture DNA modified with a digoxigen group and an anti-digoxin immobilized on a SiC@Ag substrate. The Ag@4MBA@DNA-biotin probe was prepared by linking the amino/biotin-modified probe DNA with the 4-mercaptobenzoic acid (4MBA) immobilized silver nanoparticles. Then, the "sandwich" structure was constructed by Ag@4MBA@DNA-biotin probe, target miRNA-106a and SiC@Ag@anti-digoxin/digoxin-DNA substrate to amplify the surface-enhanced Raman scattering signal. Finally, the streptavidin and the extra probes were alternately added into the “sandwich structure” to form the silver-biotin-streptavidin nanoaggregates and achieve secondary amplification of the detection signal. The experimental results show that the ultrasensitive detection of miRNA-106a has been implemented by using the dual surface-enhanced Raman scattering amplifications of SiC@Ag substrate and silver-biotin-streptavidin nanoaggregates, arriving an ultralow detection limit of 0.579 fmol/L, which has a potential for early diagnosis of tumors.

梁照恒, 王哲, 彭乐, 王福艳, 周骏. 基于SiC@Ag基底和银-生物素-链霉亲和素纳米聚集体双重SERS放大的miRNA-106a检测[J]. 光子学报, 2019, 48(7): 0717002. LIANG Zhao-heng, WANG Zhe, PENG Le, WANG Fu-yan, ZHOU Jun. Detection of miRNA-106a Based on Dual SERS Amplifications of SiC@Ag Substrate and Silver-biotin-streptavidin Nanoaggregates[J]. ACTA PHOTONICA SINICA, 2019, 48(7): 0717002.

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