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Issue 19, 2013
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Microfluidic barcode assay for antibody-based confirmatory diagnostics

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Abstract

Confirmatory diagnostics offer high clinical sensitivity and specificity typically by assaying multiple disease biomarkers. Employed in clinical laboratory settings, such assays confirm a positive screening diagnostic result. These important multiplexed confirmatory assays require hours to complete. To address this performance gap, we introduce a simple ‘single inlet, single outlet’ microchannel architecture with multiplexed analyte detection capability. A streptavidin-functionalized, channel-filling polyacrylamide gel in a straight glass microchannel operates as a 3D scaffold for a purely electrophoretic yet heterogeneous immunoassay. Biotin and biotinylated capture reagents are patterned in discrete regions along the axis of the microchannel resulting in a barcode-like pattern of reagents and spacers. To characterize barcode fabrication, an empirical study of patterning behaviour was conducted across a range of electromigration and binding reaction timescales. We apply the heterogeneous barcode immunoassay to detection of human antibodies against hepatitis C virus and human immunodeficiency virus antigens. Serum was electrophoresed through the barcode patterned gel, allowing capture of antibody targets. We assess assay performance across a range of Damkohler numbers. Compared to clinical immunoblots that require 4–10 h long sample incubation steps with concomitant 8–20 h total assay durations; directed electromigration and reaction in the microfluidic barcode assay leads to a 10 min sample incubation step and a 30 min total assay duration. Further, the barcode assay reports clinically relevant sensitivity (25 ng ml−1 in 2% human sera) comparable to standard HCV confirmatory diagnostics. Given the low voltage, low power and automated operation, we see the streamlined microfluidic barcode assay as a step towards rapid confirmatory diagnostics for a low-resource clinical laboratory setting.

Graphical abstract: Microfluidic barcode assay for antibody-based confirmatory diagnostics

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Supplementary files

Article information


Submitted
19 Feb 2013
Accepted
17 Jul 2013
First published
17 Jul 2013

Lab Chip, 2013,13, 3910-3920
Article type
Paper

Microfluidic barcode assay for antibody-based confirmatory diagnostics

M. K. Araz, A. A. Apori, C. M. Salisbury and A. E. Herr, Lab Chip, 2013, 13, 3910
DOI: 10.1039/C3LC50229E

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