A fluorescent signal "removal" sensor via duplex-specific nuclease-aided cleavage for miRNA detection in flow cytometry.

Abstract

Considered as the next-generation biomarkers, microRNAs play an important role in the early diagnosis of cancers. Here, we designed a fluorescent signal "removal" sensor for one-step, sensitive and specific detection of multiple microRNAs by flow cytometry (FCM). In this work, single-stranded DNA (ssDNA), working as the interlinkage, immobilized the fluorescent nanosphere (FS) onto the SiO2 microspheres surface to form the SiO2-ssDNA-FS probes. When target miRNAs integrated with SiO2-ssDNA-FS probes, the duplex-specific nuclease (DSN) could cleave the ssDNA selectively and release FS with numerous cycles to enhance the fluorescent signal attenuation of SiO2-ssDNA-FS, so as to remarkably improve the analysis sensitivity. It achieved a simple, accurate and quantitative microRNA-21 detection for clinical blood samples. Parallel multi-target detection of microRNA-21 and Let-7d was also realized by different color labeled FS. Moreover, our designed sensor was suitable for other targets' detection with the corresponding probes.

• Authors: Weipan Peng, Yiwen Huang, Qian Zhao, Guowei Lian, Minghui Chen, Jiafang Piao, Xiaoqun Gong, Jin Chang
• Journal: Colloids and surfaces. B, Biointerfaces (Colloids Surf B Biointerfaces) Vol. 185 Pg. 110570 (Jan 01 2020) ISSN: 1873-4367 [Electronic] Netherlands
• PMID: 31654888 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• DNA, Single-Stranded
• MicroRNAs
• Endonucleases

Topics

• DNA, Single-Stranded (metabolism)
• Endonucleases (metabolism)
• Feasibility Studies
• Flow Cytometry (methods)
• Fluorescence
• Humans
• MicroRNAs (analysis, blood, genetics)