基于无标记核酸适配体的凝血酶表面增强拉曼散射传感器  

Label-Free Aptamer-Based Surface Enhanced Raman Scattering Sensor for Thrombin

作　　者：邓媛 易润芳 周晓东[2] 胡继明[2] DENG Yuan;YI Run-fang;ZHOU Xiao-dong;HU Ji-ming(Institute of Scientific Research ＆ Development,Wuhan University,Wuhan 430072;College of Chemistry and Molecular Sciences,Wuhan University,Wuhan 430072)

机构地区：[1]武汉大学科学技术发展研究院,湖北武汉430072 [2]武汉大学化学与分子科学学院,湖北武汉430072

出　　处：《分析科学学报》2018年第5期577-582,共6页Journal of Analytical Science

基　　金：国家自然科学基金(No.41273093;81471696)

摘　　要：本文设计了一种基于无标记核酸适配体(Aptamer)的表面增强拉曼散射(SERS)传感器用于检测凝血酶。方法直接检测核酸适配体中腺嘌呤(A碱基)的环呼吸振动产生的SERS信号,并通过引入拉曼染料对巯基苯甲酸(MBA)作为内标来克服SERS光谱重现性不高的缺陷。Aptamer的热预处理、金纳米粒子溶液浓度、Aptamer和MBA在AuNPs上修饰顺序,以及探针制备过程中Aptamer和MBA加入量等实验条件进行了考察和优化。通过检测探针与凝血酶结合前后MBA和核酸适配体内源SERS信号的相对强度,及其与凝血酶浓度之间的变化关系实现了对凝血酶的定量检测。凝血酶检测浓度范围为100~2 000nmol·L^(-1),最低检测浓度为100nmol·L^(-1)。A label-free aptamer-based surface enhanced raman scattering（SERS）sensor for thrombin was designed.In this method,the SERS signal produced by cyclic respiration of adenine（A base）in aptamers was detected,and a Raman dye 4-mercaptobenzoic acid（MBA）was introduced as an internal standard to overcome the poor reproducibility of SERS spectra.The experiment conditions,including the thermal pretreatment of aptamer,the concentration of Au nanoparticles（AuNPs）solution,the adding order and quantity of aptamer and MBA modified onto AuNPs,have been investigated and optimized.By detecting the relative intensity of MBA and aptamer SERS signal before and after the binding of probe with thrombin and its relationship with the concentration of thrombin,we achieved the quantitative determination of thrombin over a concentration range from 100 nmol·L^（-1） to 2 000 nmol·L^（-1） with a minimum detectable concentration of 100 nmol·L^（-1）.

关 键 词：生物传感器 表面增强拉曼散射 凝血酶检测 无标记核酸适配体 

分 类 号：O657.37[理学—分析化学]