Ultrasensitive and Highly Specific Lateral Flow Assays for Point-of-Care Diagnosis

ACS Nano, Journal Year: 2021, Volume and Issue: 15(3), P. 3593 - 3611

Published: Feb. 19, 2021

Lateral flow assays (LFAs) are paper-based point-of-care (POC) diagnostic tools that widely used because of their low cost, ease use, and rapid format. Unfortunately, traditional commercial LFAs have significantly poorer sensitivities (μM) specificities than standard laboratory tests (enzyme-linked immunosorbent assay, ELISA: pM-fM; polymerase chain reaction, PCR: aM), thus limiting impact in disease control. In this Perspective, we review the evolving efforts to increase sensitivity specificity LFAs. Recent work improve through assay improvement includes optimization kinetics signal amplification by either reader systems or additional reagents. Together, these produced with ELISA-level (pM-fM). addition, sample preamplification can be applied both nucleic acids (direct amplification) other analytes (indirect prior LFA testing, which lead PCR-level (aM) sensitivity. However, strategies also detection time complexity, inhibits large-scale POC use Perspectives achieve future (<30 min), ultrasensitive (PCR-level), "sample-to-answer" diagnostics provided. case specificity, recent research focused on high-affinity molecules reduce nonspecific binding. Furthermore, novel highly specific molecules, such as CRISPR/Cas systems, integrated into diagnosis produce not only but diagnostics. summary, continuing improvements, may soon offer performance at is competitive techniques while retaining a

Language: Английский

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Emerging ELISA derived technologies for in vitro diagnostics

TrAC Trends in Analytical Chemistry, Journal Year: 2022, Volume and Issue: 152, P. 116605 - 116605

Published: March 25, 2022

Language: Английский

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Digital colloid-enhanced Raman spectroscopy by single-molecule counting

Nature, Journal Year: 2024, Volume and Issue: 628(8009), P. 771 - 775

Published: April 17, 2024

Language: Английский

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Framework for rapid comparison of extracellular vesicle isolation methods

eLife, Journal Year: 2021, Volume and Issue: 10

Published: Nov. 16, 2021

Extracellular vesicles (EVs) are released by all cells into biofluids and hold great promise as reservoirs of disease biomarkers. One the main challenges in studying EVs is a lack methods to quantify that sensitive enough can differentiate from similarly sized lipoproteins protein aggregates. We demonstrate use ultrasensitive, single-molecule array (Simoa) assays for quantification using three widely expressed transmembrane proteins: tetraspanins CD9, CD63, CD81. Using Simoa measure these EV markers, well albumin contamination, we were able compare relative efficiency purity several commonly used isolation plasma cerebrospinal fluid (CSF): ultracentrifugation, precipitation, size exclusion chromatography (SEC). further assays, on one platform, improve SEC CSF. Our results highlight utility quantifying proteins provide rapid framework comparing improving biofluids.

Language: Английский

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An In Situ Generated Prussian Blue Nanoparticle-Mediated Multimode Nanozyme-Linked Immunosorbent Assay for the Detection of Aflatoxin B1

ACS Applied Materials & Interfaces, Journal Year: 2021, Volume and Issue: 13(22), P. 25738 - 25747

Published: May 27, 2021

This work aims to develop a novel multimode (photothermal/colorimetric/fluorescent) nanozyme-linked immunosorbent assay (NLISA) based on the in situ generation of Prussian blue nanoparticles (PBNPs) surface magnetic (MNPs). Being considered most toxic among mycotoxins, aflatoxin B1 (AFB1) was chosen as proof-of-concept target. In this strategy, MNPs, which an AFB1 aptamer previously assembled via streptavidin-biotin linkage, are anchored 96-well plates by and antibody. presence HCl K4Fe(CN)6, PBNPs formed MNP surface, thereby achieving photothermal colorimetric signal readout due their effect intrinsic peroxidase-like activity. Based fluorescence quenching Cy5 recovered facilitate ultrasensitive detection. Photothermal signals allow portable/visual point-of-care testing, fluorescent enable accurate determination with detection limit 0.54 fg/mL, is 6333 28 times lower than those analyses, respectively. We expect that proposed NLISA can not only reduce false-positive/negative rates through multisignal crossdetection monitoring but also provide universal way sophisticated instrumentation-free, easy-to-use, cost-effective, highly sensitive other food hazards.

Language: Английский

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High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity

ACS Nano, Journal Year: 2022, Volume and Issue: 16(1), P. 1025 - 1035

Published: Jan. 14, 2022

A major challenge in many clinical diagnostic applications is the measurement of low-abundance proteins and other biomolecules biological fluids. Digital technologies such as digital enzyme-linked immunosorbent assay (ELISA) have enabled 1000-fold increases sensitivity over conventional protein detection methods. However, current ELISA still possess insufficient sensitivities for rare biomarkers require specialized instrumentation or time-consuming workflows that limited their widespread implementation. To address these challenges, we developed a more sensitive streamlined platform, Molecular On-bead Signal Amplification Individual Counting (MOSAIC), which attains low attomolar limits detection, with an order magnitude enhancement MOSAIC uses rapid, automatable flow cytometric readout vastly throughput easily integrated into existing laboratory infrastructure. As provides high sampling efficiencies target molecules, bead number can readily be tuned to enhance signal-to-background precision. Furthermore, solution-based signal expands analytes simultaneously measured higher-order multiplexing femtomolar below, compared microwell- droplet-based proof principle, apply toward improving detectability cytokines saliva ultrasensitive multiplexed measurements eight plasma saliva. The sensitivity, throughput, broad abilities provide highly accessible versatile capabilities potentially accelerate biomarker discovery testing diverse disease applications.

Language: Английский

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Development and future of droplet microfluidics

Lab on a Chip, Journal Year: 2024, Volume and Issue: 24(5), P. 1135 - 1153

Published: Jan. 1, 2024

This review introduces the development of droplet microfluidics by explaining physical mechanisms generation, discussing various approaches in manipulating droplets, and summarizing key applications material science biological analyses.

Language: Английский

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Recent advances in cascade isothermal amplification techniques for ultra-sensitive nucleic acid detection

Talanta, Journal Year: 2023, Volume and Issue: 260, P. 124645 - 124645

Published: May 4, 2023

Language: Английский

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Ultrasensitive Detection of Attomolar Protein Concentrations by Dropcast Single Molecule Assays

Journal of the American Chemical Society, Journal Year: 2020, Volume and Issue: 142(28), P. 12314 - 12323

Published: June 30, 2020

Measurements of very low levels biomolecules, including proteins and nucleic acids, remain a critical challenge in many clinical diagnostic applications due to insufficient sensitivity. While digital measurement methods such as Single Molecule Arrays (Simoa), or ELISA, have made significant advances sensitivity, there are still potential disease biomarkers that exist accessible biofluids at below the detection limits these techniques. To overcome this barrier, we developed simple strategy for single molecule counting, dropcast assays (dSimoa), enables more target molecules be counted through increased sampling efficiency with simpler workflow. In approach, beads simply onto microscope slide dried into monolayer film signal readout. The dSimoa platform achieves attomolar detection, an up 25-fold improvement sensitivity over Simoa, current state art ultrasensitive protein detection. Furthermore, its readout process improved cost-effectiveness compared existing bioassays, increases amenability integration point-of-care platforms. As illustration utility dSimoa, demonstrate ability measure previously undetectable Brachyury, tissue biomarker chordoma, plasma samples. With significantly enhanced simplicity, can pave way toward discovery new early diagnosis health outcomes.

Language: Английский

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AuPt nanocrystals/polydopamine supported on open-pored hollow carbon nanospheres for a dual-signaling electrochemical ratiometric immunosensor towards h-FABP detection

Sensors and Actuators B Chemical, Journal Year: 2021, Volume and Issue: 346, P. 130501 - 130501

Published: July 27, 2021

Language: Английский

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