Highly sensitive and label-free detection of DILI microRNA biomarker via target recycling and primer exchange reaction amplifications.
Highly sensitive and label-free detection of DILI microRNA biomarker via target recycling and primer exchange reaction amplifications.
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DOI：
10.1016/j.aca.2022.339521
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发表时间：
2022-01
影响因子：
6.2
通讯作者：
Xiaolong Li;Lei Liao;Bingying Jiang;Wenjiao Zhou;R. Yuan;Yun Xiang
中科院分区：
文献类型：
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作者：
Xiaolong Li;Lei Liao;Bingying Jiang;Wenjiao Zhou;R. Yuan;Yun Xiang
microRNA （miRNA）在生命过程中与疾病的发生发展密切相关，对microRNA生物标志物的敏感检测有利于疾病的早期诊断和治疗。本文通过将靶序列循环与引物交换反应（PER）耦合，构建了一种高灵敏度和非标记的方法来检测miRNA-122，这是药物性肝损伤的生物标志物。目标序列在燃料DNA的帮助下，通过支点介导的链置换反应，循环地置换DNA轨道上的两个发夹。释放的发夹进一步结合引物，触发聚合酶辅助的PER过程，产生大量g -四重体序列，然后与硫黄素T结合，显著增强其荧光，以49.4 fM的低检测限检测miRNA-122。该方法还可以实现对目标miRNA-122的高度特异性鉴别。该方法具有非标签格式、高选择性和高灵敏度等特点，可作为一种方便、通用的方法，用于早期疾病诊断的各种生物标志物的检测。
MicroRNAs (miRNA) are closely associated with the development of diseases in life processes, and the sensitive detection of microRNA biomarkers is beneficial to disease diagnosis and treatment at early stage. Herein, by coupling the target sequence recycling with the primer exchange reaction (PER), a highly sensitive and non-label approach is constructed to detect miRNA-122, the biomarker for drug-induced liver injury. The target sequence cyclically displaces two hairpins on the DNA track via toehold-mediated strand displacement reactions with the assistance of the fuel DNA. The released hairpins further bind the primer to trigger the polymerase-aided PER process for the yield of plenty of G-quadruplex sequences, which then combine with the thioflavin T to drastically enhance its fluorescence for sensing miRNA-122 with a low 49.4 fM detection limit. Highly specific discrimination of the target miRNA-122 can also be realized with the proposed method. Because of the non-label format, high selectivity and sensitivity, such a method can be a convenient and universal means for sensing various biomarkers for disease diagnosis at the early stages.