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Multiplexed Capillary-Flow Driven Immunoassay for Respiratory Illnesses.

Jeremy S Link1, John O'Donnell-Sloan2, Sierra Curdts1

  • 1Department of Chemistry, Colorado State University, Fort Collins, Colorado 80523, United States.

Analytical Chemistry
|February 28, 2024
PubMed
Summary
This summary is machine-generated.

This study introduces a novel multiplexed capillary-flow immunoassay for rapid, simultaneous detection of multiple biomarkers, addressing limitations of current diagnostic tests. The device enables sensitive and specific point-of-care diagnostics for diseases like influenza and COVID-19.

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Area of Science:

  • Biomarker Discovery
  • Microfluidics
  • Point-of-Care Diagnostics

Background:

  • Multiplexed analysis offers superior diagnostic capabilities over single-analyte detection in medical diagnostics.
  • Existing multiplexed methods like PCR and ELISA are time-consuming and costly.
  • Lateral flow assays (LFAs) are suitable for point-of-care testing but suffer from limitations such as poor sensitivity and cross-reactivity.

Purpose of the Study:

  • To develop the first multiplexed capillary-flow immunoassay device for simultaneous detection of multiple biomarkers.
  • To create a sensitive and rapid point-of-care diagnostic tool capable of distinguishing between diseases with similar symptoms.
  • To overcome the limitations of existing multiplexed diagnostic platforms.

Main Methods:

  • Development of a capillary-flow microfluidic platform enabling reagent and washing steps for two simultaneous ELISAs.
  • Spatially separated test strips for independent biomarker detection from a single sample addition.
  • Smartphone-based image analysis for quantitative data acquisition.

Main Results:

  • The device successfully detected and quantified H1N1 hemagglutinin (HA) and SARS-CoV-2 nucleocapsid protein (N-protein) with limits of detection of 840 pg/mL and 133 pg/mL, respectively.
  • Demonstrated no cross-reactivity between assays, with no increased signal on one test line due to the presence of the other target.
  • Successfully performed simultaneous detection of N-protein and human C-reactive protein (CRP), showing no interference between the two assays.

Conclusions:

  • The developed multiplexed capillary-flow immunoassay is a sensitive and rapid platform for simultaneous detection of multiple biomarkers.
  • This technology holds promise for point-of-care diagnostics, enabling differentiation between diseases with overlapping symptoms.
  • The device's ability to detect biomarkers like SARS-CoV-2 N-protein and CRP simultaneously offers potential for assessing disease severity.