分析化学2024，Vol.52Issue(5)：743-750,8.DOI:10.19756/j.issn.0253-3820.241117
基于黏性末端链置换扩增的电化学发光/电化学比率传感器用于miRNA检测
Ratiometric Electrochemiluminescence/Electrochemistry Sensor for miRNA Based on Toehold-Mediated Strand Displacement Amplification
摘要
Abstract
A simple and highly sensitive method for detection of miRNA was constructed by combining the high-efficiency thiolated-mediated strand displacement(TMSD)non-enzymatic amplification and the high-reproducibility electrochemiluminescence/electrochemistry(ECL/EC)ratio sensing technique.The EC signal generated by methylene blue(MB)modified on the electrode was used as the internal reference signal,and Ru(phen)32+was embedded in DNA nanoparticles produced by the branched hybridization chain reaction as the ECL probe.The presence of target miRNA facilitated the detachment of MB from electrode surface and the modification of ECL beacon on the electrode,resulting in a decrease of the EC signal and an enhancement of the ECL signal.The method of using the ratio of ECL signal to EC signal reduced potential interference and improved reproducibility and sensitivity of the biosensor.The ratio of ECL/EC signals exhibited linear relationship with miRNA-21 concentration in the range from 10 fmol/L to 500 pmol/L,with a detection limit of 6.5 fmol/L,and the relative standard deviation of the assay was reduced from 8.2%to 4.1%when determining 1 pmol/L target miRNA-21 compared to that using single ECL signal.This method was preliminarily applied to the detection of miRNA-21 in complex cell samples.关键词
电化学/电化学发光/比率传感器/黏性末端链置换/miRNAKey words
Electrochemistry/Electrochemiluminescence/Ratiometric biosensor/Toehold-mediated strand displacement/miRNA引用本文复制引用
金小亚,张莹,林悦,林振宇..基于黏性末端链置换扩增的电化学发光/电化学比率传感器用于miRNA检测[J].分析化学,2024,52(5):743-750,8.基金项目
国家自然科学基金项目(No.22174018)、浙江省基础公益研究计划项目(No.TGD23H030004)、温州市基础性科研项目(No.Y20220021)和浙江省卫生健康科技计划项目(No.2022KY887)资助. Supported by the National Natural Science Foundation of China(No.22174018),the Basic Public Welfare Research Project of Zhejiang Province(No.TGD23H030004),the Wenzhou Basic Scientific Research Project(No.Y20220021)and the Health Science and Technology Project of Zhejiang Province,China(No.2022KY887). （No.22174018）
