基于混合链式反应纳米支架的功能性核酸纳米材料级联滚动圈扩增用于信号增强型 miRNA let-7a 检测。

IF 5.3 2区 化学 Q1 CHEMISTRY, ANALYTICAL
Hao Jiang, Zhao Peng, Xuefei Lv, Ying Liu, Xiaoqiong Li, Yulin Deng
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引用次数: 0

摘要

本研究提出了一种基于杂交链式反应(HCR)纳米支架的新型功能核酸(FNA)纳米材料,利用级联扩增技术解决了miRND灵敏检测中的时间叠加和重复引物设计问题。利用制备的 FNA 纳米材料级联滚圆扩增(RCA)技术,通过侧流检测(LFA)对 miRNA let-7a(作为模型靶标)进行灵敏检测。在最佳条件下,所提出的 RCA-FNA-LFA 检测方法对 miRNA let-7a 的检测具有特异性和准确性,检测限为 1.07 pM,与 RCA -LFA 检测方法相比,灵敏度提高了近 20 倍。值得注意的是,HCR 纳米支架的非靶向依赖性自组装过程不会占用整个检测时间,因此比传统的级联法耗时更短。此外,所提出的检测方法无需考虑两种等温扩增技术之间的系统兼容性。对于不同 miRNA 的检测,只需改变 RCA 挂锁探针的同源臂,而 FNA 纳米材料则无需改变,这大大简化了级联扩增技术的引物设计。随着技术的进一步发展,RCA-FNA-LFA 检测方法可能会实现更高的灵敏度和更快的检测结果,从而更好地满足临床诊断的要求。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Hybrid chain reaction nanoscaffold-based functional nucleic acid nanomaterial cascaded with rolling circle amplification for signal enhanced miRNA let-7a detection

Hybrid chain reaction nanoscaffold-based functional nucleic acid nanomaterial cascaded with rolling circle amplification for signal enhanced miRNA let-7a detection

A novel functional nucleic acid (FNA) nanomaterial based on hybrid chain reaction (HCR) nanoscaffolds is proposed to solve the problem of time superposition and repeated primer design in sensitive miRND detection using cascade amplification technique. Rolling circle amplification (RCA) was cascaded with the prepared FNA nanomaterials for miRNA let-7a (as a model target) sensitive detection by lateral flow assay (LFA). Under the optimal conditions, the proposed RCA-FNA-LFA assay demonstrated the specificity and accuracy for miRNA let-7a detection with a detection limit of 1.07 pM, which increased sensitivity by nearly 20 times compared with that of RCA -LFA assay. It is worth noting that the non-target-dependent self-assembly process of HCR nanoscaffolds does not take up the whole detection time, thus, less time is taken than that of the conventional cascaded method. Moreover, the proposed assay does not need to consider the system compatibility between two kinds of isothermal amplification techniques. As for detection of different miRNAs, only the homologous arm of the padlock probe of RCA needs to be changed, while the FNA nanomaterial does not need any change, which greatly simplifies the primer design of the cascaded amplification techniques. With further development, the proposed RCA-FNA-LFA assay might achieve more sensitive and faster results to better satisfy the requirements of clinical diagnosis combing with more sensitive labels or small strip reader.

Graphical Abstract

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来源期刊
Microchimica Acta
Microchimica Acta 化学-分析化学
CiteScore
9.80
自引率
5.30%
发文量
410
审稿时长
2.7 months
期刊介绍: As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.
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