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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
Summary
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.
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.
