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Peer-reviewedCC BYAnalytical Chemistry

Dual Paper-Based Electrochemical Device for Multiplex Detection of Triple-Negative Breast Cancer miRNA Signatures

  • Alessandra Glovi1,2,3,4
  • Panagiota M. Kalligosfyri2,4
  • Antonella Miglione5,6
  • Sima Singh2,4
  • Wanda Cimmino2,4
  • stefania cocco7,8
  • Umberto Malapelle5,6
  • Antonio Giordano9,10,11,12
  • Michelino De Laurentiis7,8
  • Stefano Cinti2,10,11,4
12 affiliations
  1. 1Clinical and Translational Oncology
  2. 2Department of Pharmacy
  3. 3Scuola Superiore Meridionale (SSM)
  4. 4University of Naples Federico II
  5. 5Department of Public Health
  6. 6University Federico II of Naples
  7. 7Department of Breast and Thoracic Oncology
  8. 8Istituto Nazionale Tumori IRCCS “Fondazione G. Pascale”
  9. 9Department of Medical Biotechnologies
  10. 10Sbarro Institute for Cancer Research and Molecular Medicine, Center for Biotechnology, College of Science and Technology
  11. 11Temple University
  12. 12University of Siena
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Abstract

In this technical note, we demonstrate how the merging of paper-based screen-printing, engineered probes, and multiplexing might offer a potent tool toward the analysis of cancer-related miRNA signatures. In fact, while single miRNA detection cannot be associated with a health condition, the detection of miRNA signatures (more than one sequence) offers a more comprehensive reflection of disease biology. The two working electrodes are engineered with rationally designed capture probes modified with two different redox mediators, namely methylene blue and ferrocene; this is necessary to discriminate against different miRNAs, i.e., miRNA-21 and miRNA-101, which are dysregulated in triple-negative breast cancer. The use of ferrocene and methylene blue as redox mediators enables independent, interference-free target detection within a single electrochemical system. The analytical characterization of the dual sensor for both analytes is consistent with the single systems, enabling their simultaneous detection in a wide concentration range (0.1-1000 nM). The platform achieves clinically relevant low-nanomolar limits of detection in both PBS and untreated human serum, while demonstrating excellent repeatability with a relative standard deviation of ∼4%, a rapid assay time of 30 min and straightforward operational procedures. Importantly, the proof-of-concept analysis performed in untreated spiked serum samples demonstrated reliable detection within a complex biological matrix. These findings highlight the potential of multiplex miRNA signature analysis in biofluids and warrant further evaluation in clinically relevant samples. Overall, this fast, low-cost paper-based platform enables multiplex analysis at the point of care and can be readily expanded to larger miRNA panels, providing a promising foundation for future diagnostic development.

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Cite

BibTeXRIS
BibTeX
@article{glovi2026dual,
  title = {{Dual Paper-Based Electrochemical Device for Multiplex Detection of Triple-Negative Breast Cancer miRNA Signatures}},
  author = {Alessandra Glovi and Panagiota M. Kalligosfyri and Antonella Miglione and Sima Singh and Wanda Cimmino and stefania cocco and Umberto Malapelle and Antonio Giordano and Michelino De Laurentiis and Stefano Cinti},
  journal = {Analytical Chemistry},
  year = {2026},
  month = may,
  doi = {10.1021/acs.analchem.5c07951},
  url = {https://doi.org/10.1021/acs.analchem.5c07951},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Dual Paper-Based Electrochemical Device for Multiplex Detection of Triple-Negative Breast Cancer miRNA Signatures
AU  - Alessandra Glovi
AU  - Panagiota M. Kalligosfyri
AU  - Antonella Miglione
AU  - Sima Singh
AU  - Wanda Cimmino
AU  - stefania cocco
AU  - Umberto Malapelle
AU  - Antonio Giordano
AU  - Michelino De Laurentiis
AU  - Stefano Cinti
PY  - 2026
DA  - 2026/05/22
JO  - Analytical Chemistry
DO  - 10.1021/acs.analchem.5c07951
UR  - https://doi.org/10.1021/acs.analchem.5c07951
AB  - In this technical note, we demonstrate how the merging of paper-based screen-printing, engineered probes, and multiplexing might offer a potent tool toward the analysis of cancer-related miRNA signatures. In fact, while single miRNA detection cannot be associated with a health condition, the detection of miRNA signatures (more than one sequence) offers a more comprehensive reflection of disease biology. The two working electrodes are engineered with rationally designed capture probes modified with two different redox mediators, namely methylene blue and ferrocene; this is necessary to discriminate against different miRNAs, i.e., miRNA-21 and miRNA-101, which are dysregulated in triple-negative breast cancer. The use of ferrocene and methylene blue as redox mediators enables independent, interference-free target detection within a single electrochemical system. The analytical characterization of the dual sensor for both analytes is consistent with the single systems, enabling their simultaneous detection in a wide concentration range (0.1-1000 nM). The platform achieves clinically relevant low-nanomolar limits of detection in both PBS and untreated human serum, while demonstrating excellent repeatability with a relative standard deviation of ∼4%, a rapid assay time of 30 min and straightforward operational procedures. Importantly, the proof-of-concept analysis performed in untreated spiked serum samples demonstrated reliable detection within a complex biological matrix. These findings highlight the potential of multiplex miRNA signature analysis in biofluids and warrant further evaluation in clinically relevant samples. Overall, this fast, low-cost paper-based platform enables multiplex analysis at the point of care and can be readily expanded to larger miRNA panels, providing a promising foundation for future diagnostic development.
SN  - 0003-2700
SN  - 1520-6882
ER  -