Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran.
Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, Iran; Department of Pharmaceutical Biomaterials and Medical Biomaterials Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Clin Chim Acta. 2018 Aug;483:119-125. doi: 10.1016/j.cca.2018.04.031. Epub 2018 Apr 25.
In this study, we developed a colorimetric method which is primarily based on the limited amount of redox in response to the miRNA target analyte binding for the determination of the miRNA, which is used as a biomarker to confirm the clinical diagnosis of cancers such as breast cancer. This method is on the basis of the peroxidase-like property of DNA templated copper nanoclusters (DNA-CuNCs). In this research, it was found that the DNA (poly T) stabilized green emitting CuNCs which exhibited enzyme-like peroxidase activity. After hybridization of miRNA with DNA-CuNCs probe and duplex formation, CuNCs catalyzed the oxidation of the methylene blue (MB) substrate, which can interact easily with DNA/miR-155 heteroduplex. Applying the optimal conditions, the absorbance of MB decreased by increasing the target miRNA-155 with in a dynamic range from 1.0 pM to 10.0 nM down to a detection limit (LOD) of 0.6 pM. The DNA-CuNCs/MB complex was designed to develop a facile, cheap, and fast colorimetric assay for detection of miRNA through MB oxidation by DNA stabilized CuNCs.
在本研究中，我们开发了一种比色法，主要基于有限的氧化还原反应量，以响应 miRNA 靶分析物结合，用于测定 miRNA，miRNA 可用作生物标志物来确认乳腺癌等癌症的临床诊断。该方法基于 DNA 模板铜纳米簇（DNA-CuNCs）的过氧化物酶样特性。在这项研究中，发现了具有酶样过氧化物酶活性的 DNA（多 T）稳定的发绿光的 CuNCs。miRNA 与 DNA-CuNCs 探针杂交并形成双链体后，CuNCs 催化亚甲基蓝（MB）底物的氧化，MB 可以与 DNA/miR-155 异源双链轻松相互作用。在最佳条件下，随着目标 miRNA-155 的增加，MB 的吸光度降低，动态范围从 1.0 pM 到 10.0 nM 降低至检测限（LOD）为 0.6 pM。该 DNA-CuNCs/MB 复合物被设计用于通过 DNA 稳定的 CuNCs 氧化 MB 来开发一种简便、廉价、快速的比色测定法来检测 miRNA。
