Abstract
The integration of several controlled parameters within a single test system is experiencing increased demand. However, multiplexed test systems typically require complex manufacturing. Here, we describe a multiplexed immunochromatographic assay that incorporates a conventional nitrocellulose membrane, which is used together with microspot printing, to construct adjacent microfluidic “tracks” for multiplexed detection. The 1 mm distance between tracks allows for the detection of up to four different analytes. The following reagents are applied in separate zones: (a) gold nanoparticle conjugates with antibodies against each analyte, (b) other antibodies against each analyte, and (c) antispecies antibodies. The immersion of the test strip in the sample initiates the lateral flow, during which reagents of different specificities move along their tracks without track erosion or reagent mixing. An essential advantage of the proposed assay is its extreme rapidity (1–1.5 min compared with 10 min for common test strips). This assay format was applied to the detection of cardiac and inflammatory markers (myoglobin, D-dimer, and C-reactive protein) in human blood, and was characterized by high reproducibility (8%–15% coefficient of variation) with stored working ranges of conventional tests. The universal character of the proposed approach will facilitate its use for various analytes.
Highlights
The rapid progress in medical diagnostics, testing of consumer products, environmental monitoring, biosafety, and industrial control have established new requirements for the detection of biologically active compounds
This assay format was applied to the detection of cardiac and inflammatory markers in human blood, and was characterized by high reproducibility (8%–15% coefficient of variation) with stored working ranges of conventional tests
This study presents the development and characterization of a test system that implements the proposed approach for the ultra-rapid and simultaneous determination of three diagnostically important biomarkers: myoglobin (Myo), C-reactive protein (CRP), and D-dimer (DDm)
Summary
The rapid progress in medical diagnostics, testing of consumer products, environmental monitoring, biosafety, and industrial control have established new requirements for the detection of biologically active compounds. To ensure prompt and informed decision making, detection methods should provide information regarding the presence and quantity of a significant number of compounds in a minimal amount of time [1]. This trend has led to the transfer of testing from specialized laboratories to point-of-care (POC) sites, with the development of analytical devices that do not require additional tools and reagents or laborious manipulations [2]. Immunochromatographic assays ( known as lateral flow assays or test strip assays) represent the most successful approach for POC diagnostics [3,4] The devices for these assays (test strips) typically comprise multimembrane composites with preapplied reagents. The duration of the analysis is determined by the time it takes for the solution to move and for specific labeled complexes to form in sufficient quantities
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