Localized DNA tetrahedrons assisted catalytic hairpin assembly for the rapid and sensitive profiling of small extracellular vesicle-associated microRNAs

Abstract

The profiling of small extracellular vesicle-associated microRNAs (sEV-miRNAs) plays a vital role in cancer diagnosis and monitoring. However, detecting sEV-miRNAs with low expression in clinical samples remains challenging. Herein, we propose a novel electrochemical biosensor using localized DNA tetrahedron-assisted catalytic hairpin assembly (LDT-CHA) for sEV-miRNA determination. The LDT-CHA contained localized DNA tetrahedrons with CHA substrates, leveraging an efficient localized reaction to enable sensitive and rapid sEV-miRNA measurement. Based on the LDT-CHA, the proposed platform can quantitatively detect sEV-miRNA down to 25 aM in 30 min with outstanding specificity. For accurate diagnosis of gastric cancer patients, a combination of LDT-CHA and a panel of four sEV-miRNAs (sEV-miR-1246, sEV-miR-21, sEV-miR-183-5P, and sEV-miR-142-5P) was employed in a gastric cancer cohort. Compared with diagnosis with single sEV-miRNA, the proposed platform demonstrated a higher accuracy of 88.3% for early gastric tumor diagnoses with higher efficiency (AUC: 0.883) and great potential for treatment monitoring. Thus, this study provides a promising method for the bioanalysis and determination of the clinical applications of LDT-CHA. Graphical Abstract

The rate-limiting procedure of 3D DNA walkers and their applications in tandem technology

“…Inspired by nature, a spatial confinement mechanism has been established to confine dispersed reactants to a compact space to increase the chance of reactants meeting, thus accelerating the reaction efficiency. [61][62][63] To avoid this defect, Zhou et al designed a novel tetrahedral DNA framework structure for a random multipedal 3D walker (Fig. 3C).…”

Emerging Sensing and In Situ Detection Technologies for the Analysis of Extracellular Vesicle miRNAs

“…88.3% in the diagnosis of early gastric GC to the diagnosis of single EV-miRNAs. [77] HCR amplification technology is also coupled to electrochemical sensors. High amplification efficiency remains a challenge despite the fact that HCR has been used for sensitive detection of various biomarkers.…”

“…LDT‐CHA was used to detect four EV‐miRNAs associated with GC (sEV‐miR‐1246, sEV‐miR‐21 sEV‐miR‐183‐5P, and sEV‐miR‐142‐5P) with an accuracy of 88.3% in the diagnosis of early gastric GC to the diagnosis of single EV‐miRNAs. [ 77 ]…”

A sensitive and rapid electrochemical biosensor for sEV-miRNA detection based on domino-type localized catalytic hairpin assembly

“…To address these challenges, localized DNA circuits (LDCs) have been proposed [ 30 , 31 ]. LDCs use carriers with which DNA circuits are conjunct, thereby reducing the distance between reaction units and facilitating a higher regional concentration of the substrate [ 32 , 33 ].…”