Zhang Pu, Wu Xiaoyan, Chai Yaqin, Yuan Ruo
Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, People's Republic of China.
Analyst. 2014 Jun 7;139(11):2748-53. doi: 10.1039/c4an00284a.
In this study, a new universal biosensor based on luminol anodic electrochemiluminescence (ECL) for the detection of microRNA-155 was constructed by using hydrogen peroxide (H2O2) as a co-reactant and hemin as a catalyzer for signal amplification. The bare glassy carbon electrode (GCE) was first electrodeposited with Au nanoparticles (AuNPs). Then, helper DNA, which was partly complementary with the hairpin DNA chains, was assembled on the prepared GCE. Target microRNA-155 and the hairpin hybridization chains could create a formation of extended double-stranded DNA (dsDNA) polymers through the displacement of hybridization chains and the hybridization chain reaction (HCR). The HCR-generated dsDNA polymers give rise to the intercalation of a lot of hemin which could catalyze the oxidation of H2O2, leading to a remarkably amplified ECL signal output. The proposed biosensor showed a wide linear range from 5 fM to 50 pM with a relatively low detection limit of 1.67 fM for microRNA-155 detection. With excellent selectivity, good stability and high sensitivity, the proposed biosensor is promising in the development of a high-throughput assay of microRNA-155.
在本研究中,构建了一种基于鲁米诺阳极电化学发光(ECL)的新型通用生物传感器,用于检测微小RNA-155,该传感器以过氧化氢(H2O2)作为共反应剂,以血红素作为信号放大的催化剂。首先在裸玻碳电极(GCE)上电沉积金纳米颗粒(AuNPs)。然后,将与发夹DNA链部分互补的辅助DNA组装在制备好的GCE上。目标微小RNA-155与发夹杂交链可通过杂交链的置换和杂交链反应(HCR)形成延伸的双链DNA(dsDNA)聚合物。HCR产生的dsDNA聚合物会导致大量血红素嵌入,血红素可催化H2O2的氧化,从而产生显著放大的ECL信号输出。所提出的生物传感器在检测微小RNA-155时显示出5 fM至50 pM的宽线性范围,检测限相对较低,为1.67 fM。该生物传感器具有出色的选择性、良好的稳定性和高灵敏度,在开发微小RNA-155的高通量检测方法方面具有广阔前景。