双重信号放大高灵敏检测凝血酶方法研究  

Study on a new highly sensitive method for detection of thrombin based on double signal amplification

作　　者：文艳清[1] 龙倩[1] 张友玉[2] 李海涛[2] WEN Yan-qing;LONG Qian;ZHANG You-yu;LI Hai-tao(Hunan Vocational Institute of Safety Technology,Changsha 410151,China;College of Chemistry and Chemical Engineering,Hunan Normal University,Changsha 410081,China)

机构地区：[1]湖南安全技术职业学院,湖南长沙410151 [2]湖南师范大学化学化工学院,湖南长沙410181

出　　处：《化学研究与应用》2023年第5期1124-1131,共8页Chemical Research and Application

基　　金：国家自然科学基金项目(21275051,21475043)资助;湖南师范大学化学生物学及中药分析教育部重点实验室开放基金课题(KLCBTCMR2015-04)资助;湖南师范大学化学生物学及中药分析教育部重点实验室开放基金课题(KLCBTCMR18-09)资助。

摘　　要：本文介绍了一种基于无标记双重信号放大的简单、超灵敏的配体传感器,采用电化学阻抗法检测凝血酶,用氧化还原探针[Fe(CN)_(6)]^(3-/4-)分析表征电极表面电荷传递阻抗的变化(ΔR et),凝血酶与其适配体特异性结合,形成的“三明治”夹心结构,以及双链DNA刚性结构的增敏作用,可实现双重放大信号的作用,提高目标物的测定灵敏度。结果显示,线性范围良好,为0.02~10 nmol·L^(-1),检出限低,达0.22 pmol·L^(-1),方法简单、灵敏、特异性高,能用于人血清样品中凝血酶的检测。In this paper,a simple and ultra-sensitive thrombin aptasensor based on unlabeled double signal amplification is introduced.Thrombin was detected by electrochemical impedance method,and the change of electron transfer resistance(ΔR et)on the electrode surface is characterized by REDOX probe[Fe(CN)_(6)]^(3-/4-).The sandwich structure formed by the specific binding of thrombin to its aptamer and the sensitization effect of the rigid structure of double-stranded DNA can achieve the function of double amplification signal and improve the sensitivity of the target determination.The linear range of the method was 0.02~10 nmol·L^(-1),and the detection limit was 0.22 pmol·L^(-1).The method was simple,sensitive and specific,and could be used for the detection of actual samples.

关 键 词：适配体 凝血酶 双重信号放大 电化学阻抗谱(EIS) 

分 类 号：O657.1[理学—分析化学]