Cai Sheng, Ye Jiawei, Al-Maskri Abdu Ahmed Abdullah, Sun Lianli, Zeng Su
Institute of Drug Metabolism and Pharmaceutical Analysis, Zhejiang Province Key Laboratory of Anti-Cancer Drug Research, Zhejiang University, Hangzhou, Zhejiang, China.
Luminescence. 2019 Dec;34(8):823-829. doi: 10.1002/bio.3677. Epub 2019 Jul 9.
A simple microRNA (miRNA) aptasensor has been developed combining the conformational switch of a streptavidin aptamer and isothermal strand displacement amplification. In the presence of its target miRNA, the allosteric molecular beacon (aMB) probe immobilized on the plate can be 'switched on' and release the streptavidin aptamer. At the same time, Klenow fragment (3'→5' exo-) is utilized to initiate DNA-strand displacement, which starts the target recycling process. Based on the aptamer' high binding affinity and subsequent catalytic chemiluminescence (CL) detection, this CL strategy is highly specific in distinguishing mature miRNAs in same family. It exhibits a dynamic range of four orders of magnitude with a detection limit of 50 fM, and shows great potential for miRNA-related clinical practices and biochemical research.
结合链霉亲和素适体的构象转换和等温链置换扩增技术，开发了一种简单的微小RNA（miRNA）适体传感器。在其靶标miRNA存在的情况下，固定在平板上的变构分子信标（aMB）探针可以被“开启”并释放链霉亲和素适体。同时，利用klenow片段（3'→5'外切酶活性）启动DNA链置换，从而开启靶标循环过程。基于适体的高结合亲和力和随后的催化化学发光（CL）检测，这种CL策略在区分同一家族中的成熟miRNA方面具有高度特异性。它具有四个数量级的动态范围，检测限为50 fM，在与miRNA相关的临床实践和生化研究中显示出巨大潜力。
