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A highly sensitive colorimetric DNA sensor for MicroRNA-155 detection: leveraging the peroxidase-like activity of copper nanoparticles in a double amplification strategy

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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».

Funding

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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Authors and Affiliations

  1. Faculty of Sciences and Techniques, Laboratory of Process Engineering and Environment, Chemical Analysis and Biosensors Group, Hassan II University of Casablanca, P.A 146, Mohammedia, Morocco

    Maliana EL Aamri, Hasna Mohammadi & Aziz Amine

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  1. Maliana EL Aamri
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  2. Hasna Mohammadi
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  3. Aziz Amine
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Contributions

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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Correspondence to Aziz Amine.

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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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