Quantification of Multiplex miRNAs by Mass Spectrometry with Duplex-Specific Nuclease-Mediated Amplification

Early quantification of multiplex biomarkers such as microRNAs (miRNAs) is critical during disease pathologic development and therapy. To tackle challenges of low abundance and multiplexing, we herein report a mass-encoded biosensing approach with duplex-specific nuclease (DSN) mediated signal ampli...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2023-08, Vol.95 (31), p.11578-11582
Main Authors: Qian, Qing, Tang, Yuguo, Miao, Peng
Format: Article
Language:English
Subjects:
Amplification
Biomarkers
Biosensing Techniques - methods
Biosensors
Chemistry
Deoxyribonucleic acid
DNA
Enzymatic activity
Enzyme activity
Gold
Human performance
Immobilization
Iron oxides
Mass spectrometry
Mass Spectrometry - methods
Mass spectroscopy
Medical screening
Metal Nanoparticles
MicroRNAs
MicroRNAs - chemistry
miRNA
Multiplexing
Nanoparticles
Nuclease
Nucleic Acid Amplification Techniques - methods
Ribonucleases - metabolism
Scientific imaging
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Summary:Early quantification of multiplex biomarkers such as microRNAs (miRNAs) is critical during disease pathologic development and therapy. To tackle challenges of low abundance and multiplexing, we herein report a mass-encoded biosensing approach with duplex-specific nuclease (DSN) mediated signal amplification. Magnetic Fe3O4 cores are coated with small gold nanoparticles (AuNPs), which are applied to achieve facile DNA immobilization subsequent separation. This biosensor integrates multiple mass reporters corresponding to different targets (five miRNAs as examples). Due to the excellent resolution of mass spectrometry, these targets can be successfully distinguished in a single spectrum. Wide detection ranges from 10 fM to 1 nM are achieved, and the limits of detection are estimated to be 10 fM. High selectivity is promised due to the enzyme activity of DSN, and practical application in human serum samples performs satisfactorily. The number of targets to be tested can be further expanded by designing different specific mass tags in theory. Therefore, the proposed method can be utilized as an important and valuable tool to quantify multiplex miRNAs for disease screening as well as biomedical investigations.
ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.3c02541