Faculty of Materials and Chemical Engineering-Yibin University, Yibin, 64400, China.
Shanghai Pudong Hospital, Fudan University Pudong Medical Center, 2800 Gongwei Road, Pudong, Shanghai, 201399, China.
Talanta. 2022 Nov 1;249:123618. doi: 10.1016/j.talanta.2022.123618. Epub 2022 May 27.
Based on self-assembly amplification, we designed a novel microRNA (miRNA)-detection method with high specificity and sensitivity. Two unique DNA probes named Linker A and Linker B were modified with phosphorothioate (PS) at both ends. In the presence of the target miRNA, these two DNA probes were ligated together by T4 DNA ligase enzyme to form a dumbbell-shaped DNA. Then the dumbbell-shaped structure would be extended with Bst 2.0 DNA polymerase enzyme, triggering the strand displacement activity without using additional primers. These results revealed our method's ultralow detection limit (300 fM), excellent selectivity, simple operation, and capability to discriminate single-base mismatches. It is believed that this proposed approach would have great application potential in clinical diagnosis and other involved fields.
基于自组装扩增，我们设计了一种新型 miRNA（microRNA）检测方法，具有高特异性和灵敏度。两条独特的 DNA 探针分别命名为 Linker A 和 Linker B，其两端均经过硫代磷酸化（PS）修饰。在目标 miRNA 的存在下，这两条 DNA 探针通过 T4 DNA 连接酶连接在一起，形成哑铃状 DNA。然后，哑铃状结构将被 Bst 2.0 DNA 聚合酶酶延伸，在不使用额外引物的情况下触发链置换活性。这些结果表明，我们的方法具有超低的检测限（300 fM）、出色的选择性、简单的操作以及区分单碱基错配的能力。相信这种方法在临床诊断和其他相关领域具有巨大的应用潜力。
