Citation:

Detecting mir-155-3p through a Molecular Beacon Bead-Based Assay, Moreira, David, Alexandre Daniela, Miranda André, c}o Pedro Louren{\c, Baptista {Pedro V. }, Tomaz Cândida, Lu Yi, and Cruz Carla , Molecules, Volume 29, Number 13, (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 time-consuming 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.

Notes:

Funding Information: This study was developed within the scope of the CICS-UBI projects 10.54499/UIDB/00709/2020 and 10.54499/UIDP/00709/2020; LA/P/0140/2020, financed by national funds through the Portuguese Foundation for Science and Technology/MCTES; “Bolsa de Investiga{\c c}ão em Oncologia Dr. Rocha Alves do Núcleo Regional do Centro da Liga Portuguesa Contra o Cancro”, Instruct-ERIC Pilot R&D application ID 2473, and PAPILOMA ref. CENTRO-01-0145-FEDER-181235. David Moreira, André Miranda, and Daniela Alexandre acknowledge doctoral fellowship grants from FCT https://doi.org/10.54499/2023.02001.BD, https://doi.org/10.54499/2021.07695.BD and https://sciproj.ptcris.pt/80505DFA, respectively. Pedro Louren{\c c}o acknowledges a fellowship grant financed by FCT to CICS-UBI (ref. UIDP/00709/2020). Publisher Copyright: © 2024 by the authors.