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Related Experiment Video

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A Multiplex Digital Polymerase Chain Reaction-Based Platform with a Unified Readout for Multiclass Biomarker

Jiumei Hu1, Joon Soo Park2, Hanran Lei2

  • 1Department of Mechanical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, United States.

ACS Nano
|April 21, 2026
PubMed
Summary

This study introduces FluoMag-dCoDe, a novel platform for simultaneous blood biomarker quantification. It enables high-dimensional analysis of mRNA and protein biomarkers from small blood volumes, enhancing precision diagnostics.

Keywords:
digital PCRfluorescence-codinglow-volume bloodmRNA-protein codetectionmultiplex biomarker detection

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Area of Science:

  • Biotechnology
  • Molecular Diagnostics
  • Biomarker Discovery

Background:

  • Current multi-biomarker quantification methods face limitations in multiplexing capacity, workflow complexity, and data integration.
  • Accurate measurement of diverse biomarkers from blood is crucial for advancing diagnostic capabilities.

Purpose of the Study:

  • To develop a novel platform for simultaneous quantification of multiple biomarker classes from blood.
  • To overcome the limitations of existing methods for high-dimensional biomarker analysis.

Main Methods:

  • Developed FluoMag-dCoDe, a platform using fluorescence-coded magnetic beads for digital co-detection of analytes.
  • Converted diverse analytes into distinct fluorescence-coded DNA templates for multiplexed quantification in digital polymerase chain reaction (dPCR).
  • Utilized a universal primer and TaqMan probe set for simultaneous detection of eight mRNA and three protein biomarkers.

Main Results:

  • FluoMag-dCoDe achieved high quantitative fidelity with minimal cross-interference in a two-color dPCR format.
  • Demonstrated strong concordance with RT-qPCR for mRNA quantification and superior sensitivity to ELISA for protein detection.
  • Successfully quantified biomarkers from approximately 35 μL of fingerstick-compatible blood volumes.

Conclusions:

  • FluoMag-dCoDe unifies mRNA and protein quantification into a single, standardized readout.
  • The platform sustains robust performance from low blood volumes, establishing a scalable approach for high-dimensional biomarker analysis.
  • This technology advances precision diagnostics by enabling comprehensive biomarker profiling.