Chinnappan, R.; Ramadan, Q.; Zourob, M. Isolation and Detection of Exosomal mir210 Using Carbon Nanomaterial-Coated Magnetic Beads. J. Funct. Biomater.2023, 14, 441.
Chinnappan, R.; Ramadan, Q.; Zourob, M. Isolation and Detection of Exosomal mir210 Using Carbon Nanomaterial-Coated Magnetic Beads. J. Funct. Biomater. 2023, 14, 441.
Chinnappan, R.; Ramadan, Q.; Zourob, M. Isolation and Detection of Exosomal mir210 Using Carbon Nanomaterial-Coated Magnetic Beads. J. Funct. Biomater.2023, 14, 441.
Chinnappan, R.; Ramadan, Q.; Zourob, M. Isolation and Detection of Exosomal mir210 Using Carbon Nanomaterial-Coated Magnetic Beads. J. Funct. Biomater. 2023, 14, 441.
Abstract
MicroRNAs (miRNAs) within the exosomes are short non-coding RNAs that are associated with many diseases including cancers and are crucial players in regulating gene expression and regula-tion. Owing to their potential role as emerging biomarkers, efficient isolation, and quantification methods of miRNA from the exosomes in the cell’s supernatant are complicated and challenging. The quantitative PCR method is the gold standard for miRNA identification and estimation. Nucleic acid-based assays are expensive, labour-intensive, and require expertise. In this work, we demon-strated an aptamer-based magnetic separation of the exosomes, and quantification of the miRNA using a fluorescence switching assay. The fluorescence assay relies on the fluorescence of the fluorophore labelled cDNA quenched by carbon nanomaterials coated on magnetic beads (OFF state). When the target miRNA210 is introduced to the system, the cDNA detaches from the quenching surface. As a result, the fluorescence intensity increases (ON state) with increasing miRNA concentration within the dynamic range of 0-100 nM. This method can detect miR210 as low as 5 pM. This method does not exhibit any cross-reactivity with other closely related miRNAs. The method was validated by spiking miRNA with the standard RT-PCR method using SYBR green as a fluorescent probe. This method can be used for the minimally invasive detection of cancer biomarkers in breast cancer patients.
Keywords
MicroRNA detection; biosensors; pre-concentration of exosomes; breast cancer
Subject
Biology and Life Sciences, Biology and Biotechnology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
