Key Laboratory of Luminescence Analysis and Molecular Sensing Southwest University, Ministry of Education, College of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, PR China.
Henan Joint International Research Laboratory of Green Construction of Functional Molecules and Their Bioanalytical Applications, College of Chemistry, Zhengzhou University, Zhengzhou, 450001, PR China.
Anal Chim Acta. 2022 Apr 29;1204:339663. doi: 10.1016/j.aca.2022.339663. Epub 2022 Mar 15.
Herein, a mismatch-fueled catalytic hairpin assembly (MCHA) was rationally engineered, which possessed higher amplification efficiency and faster rate than catalytic hairpin assembly (CHA). Once input target microRNA-21(miRNA-21) triggers the MCHA, the hairpin DNA H1 will be opened to form the duplex H1-miRNA-21, then the mismatched hairpin DNA H2 could easily hybridize with H1-miRNA-21 to generate duplex H1-H2 and the miRNA-21 could be released to enter next cycle, thus generating amounts of output products. Impressively, the MCHA realizes a pretty shorter complete reaction time of 40 min and quite higher amplification efficiency of 9.56 × 10, which dramatically transcended the barrier: low amplification times and long reaction time in traditional CHA. As a proof of the concept, the elaborated MCHA as a hyper-efficiency and high-speed DNA signal-magnifier was successfully applied in ultrasensitive and rapid detection of miRNA-21 with the detection limit of 0.17 fM, which exploited an ingenious nucleic acid signal amplification technique for sensitive and fast detection of biomarkers in biosensing assay and clinic diagnose.
在此，理性设计了一种错配燃料催化发夹组装（MCHA），其具有比催化发夹组装（CHA）更高的扩增效率和更快的速度。一旦输入目标 microRNA-21（miRNA-21）触发 MCHA，发夹 DNA H1 将被打开形成双链 H1-miRNA-21，然后不匹配的发夹 DNA H2 可以轻松与 H1-miRNA-21 杂交生成双链 H1-H2，并且 miRNA-21 可以释放进入下一个循环，从而产生大量的输出产物。令人印象深刻的是，MCHA 实现了相当短的完整反应时间 40 分钟和相当高的扩增效率 9.56×10，这极大地突破了传统 CHA 的障碍：低扩增时间和长反应时间。作为概念验证，精心设计的 MCHA 作为超高效和高速 DNA 信号放大器，成功应用于 miRNA-21 的超灵敏和快速检测，检测限为 0.17 fM，利用巧妙的核酸信号扩增技术在生物传感分析和临床诊断中实现了生物标志物的灵敏快速检测。
