微RNA（miRNA）是一类能在癌症和其他疾病的诊断/预后生物标志物和治疗靶点中发挥重要作用的小RNA分子。本文提出了一种用于高灵敏和多功能miRNA检测的识别-切割-扩增（ICA）策略，并成功应用于miR-155和miR-21的检测。它结合了一个基于配对的识别切割和链置换扩增（ATMC-SDA）来实现ICA过程。在识别过程中，DNA配对物（DA）和DNA扩增物（DM）在靶miRNA的帮助下结合，形成一个T字结构。然后，一个结合在DA的叶片部分的识别序列上的切割核酸酶（NEase）可以在DM上进行切割，被切割的DM（CDM）可以作为引发SDA反应进行信号放大的起始子。所提出的ATMC-SDA与生物样品兼容性的测试结果显示，miR-155和miR-21的检测限度（LOD）分别为5.4aM和6.8aM，动态范围从10.0aM到10.0pM。该策略与酶和引物的组合相同，可以作为一种高灵敏检测各种miRNA的多功能ICA策略，无需反转录。同时，在人血清中的生物样品兼容性也进行了测试，结果显示具有广泛应用潜力。Copyright © 2023 Elsevier B.V. 版权所有。

MicroRNAs (miRNAs) are small RNA molecules that can play important roles as diagnostic/prognostic biomarkers and therapeutic targets for cancers and other diseases. Herein, an identification-cleavage-amplification (ICA) strategy for highly sensitive and versatile detection of miRNA has been proposed, and successfully applied to miR-155 and miR-21 assays. It combines an aligner-target mediated cleavage with strand displacement amplification (ATMC-SDA) to achieve the ICA process. During the identification process, a DNA-aligner (DA) and a DNA-amplicon (DM) can bind together with the help of target miRNA, forming a T-junction structure. Then, a nicking endonuclease (NEase), binding on the recognition sequence at the stem part of DA, can make a cleavage on DM, and the cleaved DM (CDM) can serve as an initiator to trigger the SDA reaction for signal amplification. Sharing the same set of enzymes and primers, the proposed ATMC-SDA can serve as a versatile ICA strategy for highly sensitive detection of various miRNAs, without the requirement of reverse transcription. Results show that the limits of detection (LOD) for miR-155 and miR-21 are 5.4 aM and 6.8 aM, respectively, with a dynamic range from 10.0 aM to 10.0 pM. The compatibility of ATMC-SDA with biological samples has also been tested by using human serum, indicating a promising potential for a wide variety of applications.Copyright © 2023 Elsevier B.V. All rights reserved.