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Detecting mir-155-3p through a Molecular Beacon Bead-Based Assay.

Authors :
Moreira, David
Alexandre, Daniela
Miranda, André
Lourenço, Pedro
Baptista, Pedro V.
Tomaz, Cândida
Yi Lu
Cruz, Carla
Source :
Molecules; Jul2024, Vol. 29 Issue 13, p1-10, 10p, 2 Diagrams, 3 Charts, 3 Graphs
Publication Year :
2024

Abstract

Lung cancer (LC) is recognized as one of the most prevalent and lethal cancers worldwide, underscoring an urgent need for innovative diagnostic and therapeutic approaches. MicroRNAs (miRNAs) have emerged as promising biomarkers for several diseases and their progression, such as LC. However, traditional methods for detecting and quantifying miRNAs, such as PCR, are timeconsuming and expensive. Herein, we used a molecular beacon (MB) bead-based assay immobilized in a microfluidic device to detect miR-155-3p, which is frequently overexpressed in LC. The assay relies on the fluorescence enhancement of the MB upon binding to the target miRNA via Watson and Crick complementarity, resulting in a conformational change from a stem–loop to a linear structure, thereby bringing apart the fluorophores at each end. This assay was performed on a microfluidic platform enabling rapid and straightforward target detection. We successfully detected miR-155-3p in a saline solution, obtaining a limit of detection (LOD) of 42 nM. Furthermore, we evaluated the method’s performance in more complex biological samples, including A549 cells’ total RNA and peripheral blood mononuclear cells (PBMCs) spiked with the target miRNA. We achieved satisfactory recovery rates, especially in A549 cells’ total RNA. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
14203049
Volume :
29
Issue :
13
Database :
Complementary Index
Journal :
Molecules
Publication Type :
Academic Journal
Accession number :
179361269
Full Text :
https://doi.org/10.3390/molecules29133182