Dimeric-molecular beacon based intramolecular strand displacement amplification enables robust analysis of miRNA.

Autor: Xue G; Department of Clinical Laboratory, Jiujiang City Key Laboratory of Cell Therapy, Jiujiang NO.1 People's Hospital, Jiujiang, 332000, China. Electronic address: xueguohui0816@126.com., Sui Z; College of Biological, Chemical Science and Engineering, Jiaxing University, Jiaxing, 314001, China., Chen B; College of Biological, Chemical Science and Engineering, Jiaxing University, Jiaxing, 314001, China., Xiao Z; School of Chemistry and Material Science, Huaibei Normal University, Huaibei, 235000, China. Electronic address: zbxiao@vip.henu.edu.cn., Yao Y; College of Biological, Chemical Science and Engineering, Jiaxing University, Jiaxing, 314001, China., Hua L; Department of Clinical Laboratory, Jiujiang City Key Laboratory of Cell Therapy, Jiujiang NO.1 People's Hospital, Jiujiang, 332000, China., Xu J; College of Biological, Chemical Science and Engineering, Jiaxing University, Jiaxing, 314001, China. Electronic address: jgxu0816@163.com.
Jazyk: angličtina
Zdroj: Talanta [Talanta] 2024 Dec 01; Vol. 280, pp. 126778. Date of Electronic Publication: 2024 Aug 25.
DOI: 10.1016/j.talanta.2024.126778
Abstrakt: Given the critical role of miRNAs in regulating gene expression and their potential as biomarkers for various diseases, accurate and sensitive miRNA detection is essential for early diagnosis and monitoring of conditions such as cancer. In this study, we introduce a dimeric molecular beacon (Di-MB) based isothermal strand displacement amplification (ISDA) system (Di-MB-ISDA) for enhanced miRNA detection. The Di-MB system is composed of two monomeric MBs (Mono-MBs) connected by a double-stranded DNA linker with single-stranded sequences in the middle, facilitating binding with the flexible arms of the Mono-MBs. This design forms a compact, high-density structure, significantly improving biostability against nuclease degradation. In the absence of target miRNA, the Di-MB maintains its stable structure. When target miRNA is present, it binds to the stem-loop regions, causing the hairpin structure to unfold and expose the stem sequences. These sequences serve as templates for the built-in primers, triggering DNA replication through an intramolecular recognition mechanism. This spatial confinement effect accelerates the strand displacement reaction, allowing the target miRNA to initiate additional reaction cycles and amplify the detection signal. The Di-MB-ISDA system addresses key challenges such as poor biostability and limited sensitivity seen in traditional methods. By enhancing biostability and optimizing reaction conditions, this system demonstrates robust performance for miRNA detection with a detection limit of 100 pM. The findings highlight the potential of Di-MB-ISDA for sensitive and accurate miRNA analysis, paving the way for its application in biomedical study and disease diagnosis in complex biological samples.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
Externí odkaz: