A sensing system constructed by combining a structure-switchable molecular beacon with nicking-enhanced rolling circle amplification for highly sensitive miRNA detection. Issue 9 (7th April 2022)

• Record Type: Journal Article
• Authors: Sun, Shujuan; Wang, Wenqing; Hu, Xuemei; Zheng, Cheng; Xiang, Qi; Yang, Qingguo; Zhang, Jing; Shen, Zhi-Fa; Wu, Zai-Sheng
• Is Part Of: Analyst. Volume 147:Issue 9(2022)
• Page Start: 1937
• Page End: 1943
• Publication Date: 2022-04-07
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/d1an02218k
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : A novel amplification assay strategy is developed for the highly sensitive detection of miRNA-21 based on a combination of a structure-switchable molecular beacon with nicking-enhanced rolling circle amplification. Abstract : The detection of disease-related biomarkers, including microRNA (miRNA), is of crucial importance in reducing the morbidity and mortality of cancer. Thus, there is a great desire to develop an efficient and simple sensing method to fulfill the detection of miRNAs. In this study, a novel amplification assay strategy is demonstrated for the highly sensitive detection of miRNA-21 by combining a structure-switchable molecular beacon with nicking-enhanced rolling circle amplification (SMB-NRCA). A circular padlock probe (CPP) contains a target recognition sequence, two binding sites for nicking endonuclease and three hybridization sites for SMBs. miRNA-21 can hybridize with the CPP and act as polymerization primer that initiates the rolling circle amplification (RCA) reaction and two different nicking-mediated RCA processes, releasing a large amount of SMBs and leading to a significantly amplified fluorescence signal originating from the restoration of pre-quenched fluorescence via their structural switching. Via the signal amplification based on the combination of RCA, nicking and SDA, this assay system can quantitatively detect miRNA-21 in a linear change of three orders of magnitude with a detection limit of 1 pM. The assay specificity is very high so that there is no interference from coexisting miRNAs. Moreover, the sensing system possesses ideal anti-interference ability in complicated milieux such as human serum. The novel sensing strategy shows tremendous prospects for application in tumor diagnosis and clinical therapy guidance.