A portable microchip for ultrasensitive and high-throughput assay of thrombin by rolling circle amplification and hemin/G-quadruplex system

被引：69

作者：

Lin, Xuexia ^{[1
,2
]}

Chen, Qiushui ^{[2
]}

Liu, Wu ^{[2
]}

Li, Haifang ^{[2
]}

Lin, Jin-Ming ^{[2
]}

机构：

[1] Beijing Univ Chem Technol, State Key Lab Chem Resource Engn, Beijing 100029, Peoples R China

[2] Tsinghua Univ, Beijing Key Lab Microanalyt Methods & Instrumenta, Dept Chem, Beijing 100084, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2014年 / 56卷

基金：

中国国家自然科学基金;

关键词：

Microchip; Rolling circle amplification; Colorimetric assay; G-quadruplex; DNA; PROTEINS; CAPTURE;

D O I：

10.1016/j.bios.2013.12.061

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

In this work, a convenient and high-throughput colorimetric assay was developed on an aptamer-modified microchip for ultrasensitive detection of thrombin using rolling circle amplification and G-quadruplex DNAzyme. This system consisted of an aptamer-modified microchip and a secondary aptamer. The secondary aptamer contained a thrombin aptamer and a primer with a G-quadruplex circular template. RCA technology was used to improve the sensitivity by producing the multiple G-quadruplex units. To generate colorimetric signal, G-quadruplex DNAzyme was used to catalyze the H2O2-mediated oxidation of 2,2'-azinobis (3-ethylbenzothiozoline)-6-sulfonic acid. At the optimal conditions, the linear range for thrombin was 0.100-50.000 pg/mL, and the limit of detection was down to 0.083 pg/mL. Moreover, the developed method was successfully applied to detect thrombin from human plasma and serum, indicating that this approach has great potential in clinical diagnosis and medical investigation. (C) 2014 Elsevier B.V. All rights reserved.

引用

页码：71 / 76

页数：6

共 30 条

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