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Large gold nanoparticle release assay for attomolar detection of miRNA related to myelodysplastic neoplasms
Giusy Finocchiaro1, Tomáš Špringer1, Zdeněk Krejčík2
1Institute of Photonics and Electronics of the Czech Academy of Sciences, Chaberská 1014/57, 182 00, Prague, Czech Republic.
Abstract:
MicroRNAs (miRNAs) hold potential as biomarkers for numerous cancer types, including myelodysplastic neoplasms (MDS). Here, we present a highly sensitive assay based on the oligonucleotide-triggered release of gold nanoparticles (AuNPs) for the detection of hsa-miR-451a with a surface plasmon resonance biosensor. The performance of the assay is in large part determined by the size and functional coating of AuNPs. Therefore, we investigate AuNPs in a size range from 43 to 170 nm, functionalized with thiol- or biotin-terminated oligonucleotides (AuNPsSdT or AuNPsBdT). Our study reveals that 103 nm AuNPsSdT are the best option to improve the assay performance due to their high colloidal stability, a release efficiency exceeding 90%, and a sensor response enhancement factor exceeding 105. We demonstrate that in conjunction with 103 nm AuNPsSdT, the AuNP release assay can detect hsa-miR-451a at levels down to 40 aM and quantify hsa-miR-451a physiological levels in human blood plasma. Moreover, we use the assay to demonstrate a significant down-regulation of hsa-miR-451a in blood plasma of MDS patients compared to healthy individuals, suggesting the potential relevance of hsa-miR-451a as a prospective MDS biomarker.
Insights
We developed a sensitive gold nanoparticle assay for detecting microRNA-451a, a potential biomarker for myelodysplastic neoplasms (MDS). This assay can detect low levels of microRNA-451a in blood plasma, showing its promise for MDS diagnostics.
Area of Science:
- Biomarker discovery
- Nanotechnology applications in diagnostics
- Molecular diagnostics
Background:
- MicroRNAs (miRNAs) are key regulators with potential as cancer biomarkers.
- Myelodysplastic neoplasms (MDS) require sensitive diagnostic tools.
- Gold nanoparticles (AuNPs) offer versatile platforms for biosensing.
Purpose of the Study:
- To develop a highly sensitive assay for hsa-miR-451a detection.
- To optimize AuNP characteristics for enhanced assay performance.
- To evaluate hsa-miR-451a as a potential biomarker for MDS.
Main Methods:
- Oligonucleotide-triggered release of AuNPs.
- Surface plasmon resonance (SPR) biosensor.
- Investigation of AuNP size (43-170 nm) and functionalization (thiol- or biotin-terminated).
Main Results:
- 103 nm AuNPs functionalized with thiol-terminated oligonucleotides (AuNPsSdT) optimized assay performance.
- Achieved high colloidal stability, >90% release efficiency, and >105 sensor response enhancement.
- Detected hsa-miR-451a down to 40 aM and quantified levels in human blood plasma.
- Demonstrated significant hsa-miR-451a down-regulation in MDS patients compared to healthy individuals.
Conclusions:
- The developed AuNP release assay with optimized AuNPs offers high sensitivity for hsa-miR-451a detection.
- hsa-miR-451a shows potential as a diagnostic biomarker for myelodysplastic neoplasms.
- This assay platform can be applied to other miRNA biomarker studies.
