An Improved System for Quantitative Immunoassay Measurement in ImageQuant

Shah, Malay Ilesh; Rajagopalan, A. N.; Joseph, Jayaraj; Sivaprakasam, Mohanashankar

doi:10.1109/icsens.2018.8589936

Cited by 5 publications

(9 citation statements)

References 10 publications

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“…This lack of background fluorescence indicates that Whatman paper is compatible with fluorescence detection. Following image processing by the instrument inbuilt software, the pixel volumes of the test (V T ), and control (V C ) lines were calculated and then used to determine the pixel volume ratio, V R (see Shah et al 2018 for details [26]).…”

Section: Resultsmentioning

confidence: 99%

“…Ltd. (Ambala, India) was used to dispense lines over the analytical strip. The fluorescence signals that were generated at the LFA test and control lines were evaluated using ImageQuant ( Figure S1 (Supplementary Materials) ), a portable immunoanalyzer developed and designed at the Healthcare Technology Innovation Center (IIT, Madras, India) [ 25 , 26 ]. This instrument relies on a laser based confocal optics system to capture the fluorescence of the test and control lines of the LFA strips.…”

Section: Methodsmentioning

confidence: 99%

“…The captured images are analyzed with LabVIEW™ software (National Instruments, Austin, TX, USA) to obtain signal data from test and control lines. The system uses intelligent image-analytics techniques that identify the reaction kinematics from a sequence of images, track the reaction progress and development of the test and control lines, identify the stabilization of the reaction, and calculate the test and control line areas and area ratios [ 25 , 26 ].…”

Section: Methodsmentioning

confidence: 99%

“…All of the quantitative data were assessed with GraphPad Prism 6.0 (GraphPad Software, La Jolla, CA, USA). Fluorescence intensity data were used to calculate the pixel volume of the test, V T , and control, V C , lines, which correspond to the two-dimensional summation of all pixel intensities within each line [ 26 ]. The corresponding mean volume ratio, V R , defined as the ratio V T /V C , was plotted versus the cTnI concentration to generate calibration curves.…”

Section: Methodsmentioning

confidence: 99%

“…The system relies on the recognition of the target cTnI molecule by fluorescently labelled anti-cTnI detection antibodies and subsequent capture of the complex at the test line by an anti-cTnI capturing antibody ( Figure 1 b,c). The fluorescent signal that is generated at the LFA lines is then quantified while using a portable immunoanalyzer [ 25 , 26 ].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

A Cellulose Paper-Based Fluorescent Lateral Flow Immunoassay for the Quantitative Detection of Cardiac Troponin I

2021

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This paper presents a lateral flow assay (LFA) for the quantitative, fluorescence-based detection of the cardiac biomarker troponin I (cTnI) that features an analytical strip made of cellulose filter paper. The results show that the wicking and test time are comparable to those obtained with conventional nitrocellulose (NC)-based LFAs. Further, the cellulose paper provides an excellent background with no auto-fluorescence that is very adequate in detecting fluorescent lines. While fluorescence that was generated with cellulose strips was lower when compared to that generated in NC strips, signals could be improved by layering carbon nanofibers (CNF) on the cellulose. A nonlinear behavior of the concentration–response relationship was observed for the LFA architectures with NC, cellulose, and cellulose-CNF in the 0 to 200 ng/mL cTnI concentration range. The measurements were consistent and characterized by coefficients of variation lower than 2.5%. Detection and quantitation limits that were in the range 1.28–1.40 ng/mL and 2.10–2.75 ng/mL were obtained for LFA with cellulose and cellulose CNF strips that are equivalent to the limits obtained with the standard NC LFA. Overall, we showed that commercially available filter paper can be used in the analytical strip of LFA.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Cellulose Paper-Based Fluorescent Lateral Flow Immunoassay for the Quantitative Detection of Cardiac Troponin I

2021

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Exploring carbohydrate binding module fusions and Fab fragments in a cellulose-based lateral flow immunoassay for detection of cystatin C

Natarajan

Joseph

Prazeres

2022

Sci Rep

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This paper presents a lateral flow assay (LFA) for the quantitative, fluorescence-based detection of the kidney biomarker cystatin C that features conjugates of capture antibodies and fusions of carbohydrate binding modules (CBM) with ZZ domains anchored on cellulose deposited over nitrocellulose (NC). The ZZ-CBM3 fusion provides a biomolecular interface between the cellulose layer and the Fc portion of the capture antibodies. By resorting to detection Fab fragments that lack the Fc portion we overcome the observed interference of full-length detection antibodies with the ZZ-CBM3 fusion at the test lines. Using the new LFA architecture, a linear concentration–response relationship was observed in the 0–10 ng/mL cystatin C concentration range, which is compatible with the clinically normal (5–120 ng/mL) and abnormal (> 250 ng/mL) levels of cystatin C, as long as proper dilutions are made. An inter assay CoV of 0.72% was obtained. Finally, mock urine samples characteristic of normal (100 ng/mL) and kidney tubular disease (4000 ng/mL) patients were successfully analyzed. Overall, we demonstrate an innovative LFA architecture that combines NC strips with layered cellulose, ZZ-CBM3 fusions and fluorescently labeled Fab fragments.

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Aptamer based Lateral Flow Assays for Rapid and Sensitive Detection of CKD marker Cystatin C

Natarajan

DeRosa

Joseph

et al. 2021

2021 IEEE International Symposium on Medical Measurements and Applications (MeMeA)

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An Improved System for Quantitative Immunoassay Measurement in ImageQuant

Cited by 5 publications

References 10 publications

A Cellulose Paper-Based Fluorescent Lateral Flow Immunoassay for the Quantitative Detection of Cardiac Troponin I

A Cellulose Paper-Based Fluorescent Lateral Flow Immunoassay for the Quantitative Detection of Cardiac Troponin I

Exploring carbohydrate binding module fusions and Fab fragments in a cellulose-based lateral flow immunoassay for detection of cystatin C

Aptamer based Lateral Flow Assays for Rapid and Sensitive Detection of CKD marker Cystatin C

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