Abstract
A novel and highly sensitive colorimetric DNA sensor for determination of miRNA-155 at attomolar levelsis presented that combines the peroxidase-like activity of copper nanoparticles (CuNPs) with the hybridization chain reaction (HCR) . The utilization of CuNPs offers advantages such as strong interaction with double-stranded DNA, excellent molecular recognition, and mimic catalytic activity. Herein, a capture probe DNA (P1) was immobilized on carboxylated magnetic beads (MBs), allowing for amplified immobilization due to the 3D surface. Subsequently, the presence of the target microRNA-155 led to the formation of a sandwich structure (P2/microRNA-155/P1/MBs) when P2 was introduced to the modified P1/MBs. The HCR reaction was then triggered by adding H1 and H2 to create a super sandwich (H1/H2)n. Following this, Cu2+ ions were attracted to the negatively charged phosphate groups of the (H1/H2)n and reduced by ascorbic acid, resulting in the formation of CuNPs, which were embedded into the grooves of the (H1/H2)n. The peroxidase-like activity of CuNPs catalyzed the oxidation reaction of 3,3',5,5'-Tetramethylbenzidine (TMB), resulting in a distinct blue color measured at 630 nm. Under optimal conditions, the colorimetric biosensor exhibited a linear response to microRNA–155 concentrations ranging from 80 to 500 aM, with a detection limit of 22 aM, and discriminate against other microRNAs. It was also successfully applied to the determination of microRNA–155 levels in spiked human serum.
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Acknowledgements
The authors acknowledge the Moroccan Ministry of Higher Education, Scientific Research and Innovation and the OCP Foundation « APRD research program 2021».
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This research was funded by the Moroccan Ministry of Higher Education, Scientific Research and Innovation and the OCP Foundation « APRD research program 2021».
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Maliana El Aamri: Methodology, Fabrication, Data Curation, Investigation, Writing-Original draft preparation. Hasna Mohammadi: Writing-Review & Editing, Review & Editing, Supervision, funding acquisition. Aziz Amine: Writing-Review & Editing, Review & Editing, Supervision, funding acquisition.
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Highlights
• An original non-enzymatic bio-assay was developed for an attomolar-level colorimetric microRNA-155 detection.
• The ingenious combination of HCR with the peroxidase-like activity of copper nanoparticles enhances sensitivity while concurrently lowering the detection limit.
• An ultralow detection limit of 22 aM was achieved for the direct detection of microRNA-155 biomarker in human serum samples.
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EL Aamri, M., Mohammadi, H. & Amine, A. A highly sensitive colorimetric DNA sensor for MicroRNA-155 detection: leveraging the peroxidase-like activity of copper nanoparticles in a double amplification strategy. Microchim Acta 191, 32 (2024). https://doi.org/10.1007/s00604-023-06087-1
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DOI: https://doi.org/10.1007/s00604-023-06087-1