A universal colorimetry for nucleic acids and aptamer-specific ligands detection based on DNA hybridization amplification
A universal colorimetry for nucleic acids and aptamer-specific ligands detection based on DNA hybridization amplification
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基于 DNA 杂交扩增的核酸和适体特异性配体检测的通用比色法
DOI：
10.1016/j.ab.2017.04.013
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发表时间：
2017-07-01
影响因子：
2.9
通讯作者：
You, Jinmao
中科院分区：
文献类型：
--
作者：
Li, Shuang;Shang, Xinxin;You, Jinmao
提出了一种基于金纳米粒子-DNA杂交链反应（HCR）的通用荧光比色法，用于检测核酸靶或适体特异性配体靶。通用芯片由捕获探针和发夹DNA-GNP组成。首先，捕获探针识别靶特异性并释放起始序列。然后分散的发夹DNA修饰的GNP通过引发序列触发的HCR事件交联形成聚集体。随着聚集体的积累，肉眼可以很容易地观察到明显的红色至紫色的颜色变化。我们使用miRNA靶序列（miRNA-203）和适体特异性配体（ATP）作为该概念验证实验的靶分子。在miRNA-203的强烈竞争下，起始序列（DNA 2）从捕获探针（MNP/DNA 1/2缀合物）中释放出来。发夹DNA（H1和H2）可以在起始DNA 2的帮助下互补以形成GNP-H1/GNP-H2聚集体。溶液的吸收比（A（620）/A（520））值是miRNA-203浓度的敏感函数，范围为1.0 × 10（-11）M至9.0 × 10（-10）M，并且可以检测到低至1.0 × 10（-11）M的miRNA-203。同时，溶液的颜色也发生了由浅酒红色到紫色再到浅蓝色的变化。对于ATP，在适体-ATP的强结合下，起始序列（DNA 3的5 '末端）从捕获探针（DNA 3）释放。该比色法具有良好的灵敏度，可检测1.0 × 10 ~（-8）M ATP。所提出的策略也表现出良好的性能，用于细胞内核酸和适体特异性配体的定性分析和定量分析。(C)2017爱思唯尔公司All rights reserved.
We present a universal amplified-colorimetric for detecting nucleic acid targets or aptamer-specific ligand targets based on gold nanoparticle-DNA (GNP-DNA) hybridization chain reaction (HCR). The universal arrays consisted of capture probe and hairpin DNA-GNP. First, capture probe recognized target specificity and released the initiator sequence. Then dispersed hairpin DNA modified GNPs were cross linked to form aggregates through HCR events triggered by initiator sequence. As the aggregates accumulate, a significant red-to purple color change can be easily visualized by the naked eye. We used miRNA target sequence (miRNA-203) and aptamer-specific ligand (ATP) as target molecules for this proof-of-concept experiment. Initiator sequence (DNA2) was released from the capture probe (MNP/DNA1/2 conjugates) under the strong competitiveness of miRNA-203. Hairpin DNA (H1 and H2) can be complementary with the help of initiator DNA2 to form GNP-H1/GNP-H2 aggregates. The absorption ratio (A(620)/A(520)) values of solutions were a sensitive function of miRNA-203 concentration covering from 1.0 x 10(-11) M to 9.0 x 10(-10) M, and as low as 1.0 x 10(-11) M could be detected. At the same time, the color changed from light wine red to purple and then to light blue have occurred in the solution. For ATP, initiator sequence (5'-end of DNA3) was released from the capture probe (DNA3) under the strong combination of aptamer-ATP. The present colorimetric for specific detection of ATP exhibited good sensitivity and 1.0 x 10(-8) M ATP could be detected. The proposed strategy also showed good performances for qualitative analysis and quantitative analysis of intracellular nucleic acids and aptamer-specific ligands. (C) 2017 Elsevier Inc. All rights reserved.