Novel Biomarkers Detection and Identification by Microfluidic- Based MicroELISA

Lakkis, Montaha; Kai, Junhai; Santiago, Nelson

doi:10.4172/2161-1025.s1-006

Cited by 3 publications

(2 citation statements)

References 9 publications

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“…In this work, we describe implementation of the next generation commercial microfluidic-based microplate to perform sandwich ELISA for screening plant pathogens commonly found in cucurbits. To our knowledge, this miniaturized system has only been instrumental in human cytokine biomarker (IL4) detection [24] and cell signaling pathway study [25] . This study, thus, presents the first demonstration of this miniaturized sandwich ELISA technology for detecting seedborne pathogens: Acidovorax avenae subsp.…”

Section: Introductionmentioning

confidence: 99%

Implementation of Microfluidic Sandwich ELISA for Superior Detection of Plant Pathogens

et al. 2013

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Rapid and economical screening of plant pathogens is a high-priority need in the seed industry. Crop quality control and disease surveillance demand early and accurate detection in addition to robustness, scalability, and cost efficiency typically required for selective breeding and certification programs. Compared to conventional bench-top detection techniques routinely employed, a microfluidic-based approach offers unique benefits to address these needs simultaneously. To our knowledge, this work reports the first attempt to perform microfluidic sandwich ELISA for Acidovorax citrulli (Ac), watermelon silver mottle virus (WSMoV), and melon yellow spot virus (MYSV) screening. The immunoassay occurs on the surface of a reaction chamber represented by a microfluidic channel. The capillary force within the microchannel draws a reagent into the reaction chamber as well as facilitates assay incubation. Because the underlying pad automatically absorbs excess fluid, the only operation required is sequential loading of buffers/reagents. Buffer selection, antibody concentrations, and sample loading scheme were optimized for each pathogen. Assay optimization reveals that the 20-folds lower sample volume demanded by the microchannel structure outweighs the 2- to 4-folds higher antibody concentrations required, resulting in overall 5–10 folds of reagent savings. In addition to cutting the assay time by more than 50%, the new platform offers 65% cost savings from less reagent consumption and labor cost. Our study also shows 12.5-, 2-, and 4-fold improvement in assay sensitivity for Ac, WSMoV, and MYSV, respectively. Practical feasibility is demonstrated using 19 real plant samples. Given a standard 96-well plate format, the developed assay is compatible with commercial fluorescent plate readers and readily amendable to robotic liquid handling systems for completely hand-free assay automation.

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Section: Introductionmentioning

confidence: 99%

Implementation of Microfluidic Sandwich ELISA for Superior Detection of Plant Pathogens

et al. 2013

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“…Lab-on-chip (LOC) devices have attractive performance characteristics (specificity, sensitivity, and high accuracy) that allow them to be powerful point-of-care (POC) diagnostic tools to examine molecular interactions at levels not always readily available in traditional larger scale instrumentation. Of particular interest has been the use of LOC devices to perform ELISAs where turnaround time is faster and sample consumption is lower than traditional assay technique [2][3][4]. Furthermore, microfluidic platforms afford unique properties due to miniaturization including small volume, large surface-to-volume ratio, laminar flow, short diffusion distance, and high surface tension.…”

mentioning

confidence: 99%

Development of a microfluidic‐based assay on a novel nitrocellulose platform

et al. 2015

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A novel microfluidic paper-based analytical device (μPAD) utilizing a nitrocellulose (NC) membrane to detect IgG antibodies through a colorimetric analysis is described. The μPAD was constructed using layered polyethylene terephthalate (PET) and pressure-sensitive adhesives (PSA). The biotin labeled Goat Anti-Mouse IgG antibody was spotted and dried on the NC channel prior to subjecting it to a series of wash solutions (Tris-tween), increasing concentrations of alkaline phosphatase conjugated to streptavidin (Strep-ALP), and para-nitrophenyl phosphate (p-NPP) realizing a vibrant yellow color. The reaction proceeds for 10 min before applying the p-NPP stop solution. The device was then dried, scanned, and analyzed yielding a linear range of inverse yellow color intensities versus Strep-ALP concentrations. The development of this simple μPAD should further facilitate the use of NC in colorimetric assays to detect and quantitate antibodies.

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Performance evaluation of microfluidic microplate-based fluorescent ELISA for qualitative detection of SARS-CoV-2–specific IgG and IgM

Kweon,

Yoon,

Choe

et al. 2024

Sci Rep

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We evaluated the diagnostic performance of newly developed microfluidic microplate-based fluorescent ELISA for anti-SARS-CoV-2 antibody detection: the Veri-Q opti COVID-19 IgG and IgM ELISAs (hereafter, “Opti IgG/M”; MiCo BioMed, Gyeonggi-do, Republic of Korea), in comparison with conventional ELISAs. A total of 270 serum samples were analyzed, among which 90 samples were serially obtained from 25 COVID-19 patients. Another 180 samples were collected from 180 SARS-CoV-2–negative individuals. As comparative assays, we used SCoV-2 Detect IgG/M ELISA (hereafter, “InBios IgG/M”; InBios, Seattle, WA, USA) and Veri-Q COVID-19 IgG/IgM ELISA (hereafter, “Veri-Q IgG/M”; MiCo BioMed). Compared with conventional ELISAs, the Opti IgG yielded 97.1–100.0% positive percent agreement, 95.2–98.0% negative percent agreement, 96.3–97.8% total percent agreement, and kappa values of 0.90–0.94. Between the Opti IgM and the InBios IgM, the values were 93.7%, 96.6%, 95.9%, and 0.89, respectively. For the Opti IgG, sensitivities for the samples collected from 0–7, 8–14, 15–21, and ≥ 22 days after symptom onset were 40.0, 58.3, 94.1, and 100.0%, respectively. The values for the Opti IgM were 30.0, 54.2, 88.2, and 80%, respectively. The diagnostic specificities of the Opti IgG and IgM were 99.4 and 97.2%, respectively. The microfluidic microplate-based fluorescent ELISAs showed comparable diagnostic performance to conventional ELISAs for detecting anti-SARS-CoV-2 antibodies. With the combination of high throughput, a simplified workflow, and the ability to analyze reduced volumes, this new technology has great potential for improving SARS-CoV-2 serologic testing.

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Novel Biomarkers Detection and Identification by Microfluidic- Based MicroELISA

Cited by 3 publications

References 9 publications

Implementation of Microfluidic Sandwich ELISA for Superior Detection of Plant Pathogens

Implementation of Microfluidic Sandwich ELISA for Superior Detection of Plant Pathogens

Development of a microfluidic‐based assay on a novel nitrocellulose platform

Performance evaluation of microfluidic microplate-based fluorescent ELISA for qualitative detection of SARS-CoV-2–specific IgG and IgM

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