Paper-microfluidic signal-enhanced immunoassays

Sathishkumar, N.; Toley, Bhushan J.

doi:10.1016/bs.pmbts.2021.07.015

Cited by 6 publications

(5 citation statements)

References 69 publications

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“…Lateral Flow Assays (LFAs) have emerged as valuable paper-microfluidic tools for point-of-care (POC) diagnostics, offering a quick, economical, and user-friendly method to detect the presence of analytes in complex samples [1][2][3][4][5][6][7][8] . Primarily known for their application in medical diagnostics, environmental monitoring, and food safety, LFAs have gained significant attention for their potential in point-of-care testing, especially in resource-limited settings 2,4,7,9,10 . Accurate and reproducible quantification and interpretation of the assay results is crucial for reliable diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Streamlining Lateral Flow Assay (LFA) Data Analysis: A MATLAB® Live Script for Integrated Batch Image Processing and Limit of Detection Determination

Saxena,

Toley

2024

Preprint

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Lateral flow assays (LFAs) are widely utilized for rapid point-of-care diagnostics in various fields; however, their quantitative image analysis is frequently hindered by repetitive manual processes and inconsistencies, typically involving the use of combinations of image processing and data analysis software tools. This protocol introduces a MATLAB live-script-based code for the integrated batch processing of LFA images and standard curve development. This code simplifies quantitative LFA analysis by automating image processing, peak detection, statistical analysis, and data fitting to a 4-parameter logistic curve for accurate Limit of Detection (LoD) determination. This approach significantly minimizes manual intervention and the potential for human error, thereby enhancing reproducibility. This addresses the critical need for efficient and reliable LFA data analysis in research laboratories. By offering a standardized method for LFA image processing and data analysis, this protocol facilitates faster and more consistent research outcomes.

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Section: Introductionmentioning

confidence: 99%

Streamlining Lateral Flow Assay (LFA) Data Analysis: A MATLAB® Live Script for Integrated Batch Image Processing and Limit of Detection Determination

Saxena,

Toley

2024

Preprint

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“…This blocking reduces the binding of the label in the analytical zone and makes reliable serodiagnostics difficult at low contents of specific antibodies [16,17]. The use of additional stages of the assay to enhance the recorded signal is possible [18,19], but it deprives LFIA of its main advantages in rapidity and easy implementation. This limitation can be overcome in various ways.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Three Lateral Flow Immunoassay Formats for the Detection of Antibodies against the SARS-CoV-2 Antigen

et al. 2023

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Reliable detection of specific antibodies against pathogens by lateral flow immunoassay (LFIA) greatly depends on the composition of the detectable complex and the order of its assembly. We compared three LFIA formats for revealing anti-SARS-CoV-2 antibodies in sera with the following detected complexes in the analytical zone of the strip: antigen–antibodies–labeled immunoglobulin-binding protein (Scheme A); antigen–antibodies–labeled antigen (Scheme B); and immunoglobulin-binding protein–antibodies–labeled antigen (Scheme C). The lowest detection limit was observed for Scheme C, and was equal to 10 ng/mL of specific humanized monoclonal antibodies. When working with pooled positive sera, Scheme C had a detection limit 15 times lower than Scheme B and 255 times lower than Scheme A. Due to the high sensitivity of Scheme C, its application for the panel of human sera (n = 22) demonstrated 100% diagnostic specificity and sensitivity. These consistent results be useful for designing the format of LFIA serodiagnosis for other diseases.

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“…Lateral flow immunoassays (LFAs) have revolu onized point-of-care diagnos cs due to their simplicity, speed, and costeffec veness. [1][2][3] However, achieving high sensi vity remains a challenge, par cularly when detec ng low analyte concentra ons. Conven onal LFAs u lizing gold nanopar cles (AuNPs) as reporters have limita ons in sensi vity, o en falling short compared to more sensi ve techniques such as enzyme-linked immunosorbent assays (ELISA) or polymerase chain reac on (PCR) [1] .…”

Section: Introduc Onmentioning

confidence: 99%

“…Thus, there is a need for innova ve strategies to enhance the sensi vity of LFAs.One approach to improve LFA sensi vity is the use of enzyma c LFAs, in which the detec on an body is conjugated to an enzyme, such as horseradish peroxidase (HRP). Upon binding to the target analyte, the enzyme-labelled an body generates a detectable signal through a cataly c reac on [2][3][4] . This enzyma c amplifica on mechanism provides a means to achieve higher sensi vity in LFAs by increasing the signal output and facilita ng the detec on of low analyte concentra ons.The present work focuses on developing enzyma cally enhanced lateral flow immunoassays capable of detec ng sub-ng/mL concentra ons of an gens.…”

mentioning

confidence: 99%

Novel Oxidative Coupling-Based Chromogenic Substrates for Horseradish Peroxidase-Enhanced Lateral Flow Immunoassays: A Highly Sensitive and Economical Alternative to Conventional Substrates

Saxena

Toley

2023

Preprint

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Lateral flow immunoassays (LFIAs) have become indispensable in point-of-care diagnostics, but improving their sensitivity remains a critical challenge. Here, we present a strategy to enhance LFA sensitivity by introducing novel oxidative coupling-based chromogenic substrates for horseradish peroxidase (HRP) enhancement. Motivated by the need for more sensitive and cost-effective LFAs, we developed substrates inspired by oxidative permanent hair dyes. These substrates consist of two components: a primary intermediate and a coupler that undergo an oxidative coupling reaction in the presence of HRP and H2O2 to generate an intensely coloured dye. Our LFAs employing these substrates outperform conventional substrates (3,3'-diaminobenzidine (DAB) and 3,3',5,5'-tetramethylbenzidine (TMB)) and achieve an unprecedented sub-ng/mL limit of detection (LoD) for Human IgG. LFAs employing these novel substrates could detect Human IgG at 0.2 ng/mL, significantly surpassing reported sensitivities in the literature. Compared to gold nanoparticle based LFAs, our substrates exhibit over 30 times higher sensitivity. Moreover, these substrates offer substantial cost savings, making them a compelling choice for point-of-care diagnostics.

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Paper-microfluidic signal-enhanced immunoassays

Cited by 6 publications

References 69 publications

Streamlining Lateral Flow Assay (LFA) Data Analysis: A MATLAB® Live Script for Integrated Batch Image Processing and Limit of Detection Determination

Streamlining Lateral Flow Assay (LFA) Data Analysis: A MATLAB® Live Script for Integrated Batch Image Processing and Limit of Detection Determination

Comparison of Three Lateral Flow Immunoassay Formats for the Detection of Antibodies against the SARS-CoV-2 Antigen

Novel Oxidative Coupling-Based Chromogenic Substrates for Horseradish Peroxidase-Enhanced Lateral Flow Immunoassays: A Highly Sensitive and Economical Alternative to Conventional Substrates

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