Programmable DNA barcode-encoded exponential amplification reaction for the multiplex detection of miRNAs

作者全名："Tan, Yuqian; Zhang, Li; Deng, Shixiong"

作者地址："[Tan, Yuqian; Zhang, Li; Deng, Shixiong] Chongqing Med Univ, Dept Forens Med, Chongqing 400016, Peoples R China"

通信作者："Deng, SX (通讯作者)，Chongqing Med Univ, Dept Forens Med, Chongqing 400016, Peoples R China."

来源：ANALYTICAL METHODS

ESI学科分类：CHEMISTRY

WOS号：WOS:001174219000001

JCR分区：Q1

影响因子：2.7

年份：2024

卷号：16

期号：11

开始页：1649

结束页：1658

文献类型：Article

关键词：　

摘要："Multiple analysis of miRNAs is essential for the early diagnosis and monitoring of diseases. Here, a programmable, multiplex, and sensitive approach was developed for one-pot detection of miRNAs by melting temperature encoded sequences and exponential isothermal amplification (E-EXPAR). In the presence of target miRNAs, the corresponding templates initiate the cycles of nicking and polymerization/displacement, generating numerous barcode strands with unique encoding sequences. Subsequently, generated barcode strands hybridize with fluorescent probes and quench the fluorophore by a triplet of G base through a photo-induced electron transfer mechanism. Finally, a melting curve analysis is performed to quantify miRNAs by calculating the rate of fluorescence change at the corresponding melting temperature. Based on this, miRNA-21, miRNA-9, and miRNA-122 were detected with the detection limits of 3.3 fM, 2.9 fM, and 1.7 fM, respectively. This E-EXPAR was also employed to simultaneously detect three miRNAs in biological samples, showing consistent results with RT-qPCR. Overall, this study provides a programmable and universal platform for multiplex analysis of miRNAs, and holds great promise as an alternative to the multiplex analysis in clinical diagnostics and prognostics for nucleic acid detection. Multiple analysis of miRNAs is essential for the early diagnosis and monitoring of diseases."

基金机构：Natural Science Foundation of Chongqing Municipality [cstc2021jcyj-msxmX0485]; Natural Science Foundation of Chongqing Province

基金资助正文：This work was sponsored by the Natural Science Foundation of Chongqing Province [grant number cstc2021jcyj-msxmX0485].