Development and statistical assessment of a paper-based immunoassay for detection of tumor markers

Mazzu-Nascimento, Thiago; Morbioli, Giorgio Gianini; Milan, Luís Aparecido; Donofrio, Fabiana Cristina; Mestriner, Carlos Alberto; Carrilho, Emanuel

doi:10.1016/j.aca.2016.11.011

Cited by 43 publications

(16 citation statements)

References 44 publications

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“…There are several well studied HRP chromogenic substrates, such as: 3,3',5,5'-tetramethylbenzidine (TMB); 2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS); o-phenylenediamine (OPD); 5-aminosalicylic acid (5-AS); 3-amino-9-ethylcarbazole (AEC); 3-methyl-2-benzothiazolinone hydrazone (MBTH); 3,3'-diaminobenzidine (DAB) and 4-chloro-1-naphthol (4-CN) [16][17][18]. In addition, the detection of the colorimetric signal can be further enhanced, in order to allow a more accurate signal measurement using a selected stopping solution [19,20].…”

Section: Introductionmentioning

confidence: 99%

Enhanced Colorimetric Signal for Accurate Signal Detection in Paper-Based Biosensors

Harpaz

Eltzov

et al. 2020

Diagnostics

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Paper-based colorimetric biosensors combine the use of paper with colorimetric signal detection. However, they usually demonstrate lower sensitivities because a signal amplification procedure has not been used. Stopping the reaction of colorimetric signal generation is often used in lab-based assays in order to amplify and stabilize the colorimetric signal for detection. In this study, the generation of a stopped colorimetric signal was examined for accurate and enhanced signal detection in paper-based biosensors. The colorimetric reaction in biosensors is usually based on the interaction between the enzyme horseradish peroxidase (HRP) and a selected chromogenic substrate. The two most commonly used HRP substrates, 3,3’,5,5’-tetramethylbenzidine (TMB) and 2’-azinobis (3-ethylbenzothiazoline-6-sulfonic-acid) (ABTS), were compared in terms of their ability to generate a stopped colorimetric signal on membrane. The stopped colorimetric signal was visible for TMB but not for ABTS. Moreover, the generation of stopped colorimetric signal was dependent on the presence of polyvinylidene-difluoride (PVDF) membrane as the separation layer. With PVDF the colorimetric signal (color intensity) was higher (TMB: 126 ± 6 and ABTS: 121 ± 9) in comparison to without PVDF (TMB: 110 ± 2 and ABTS: 102 ± 4). The TMB stopped colorimetric signal demonstrated a more stable signal detection with lower standard deviation values. To conclude, a stopped colorimetric signal can be generated in paper-based biosensors for enhanced and accurate signal detection.

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Section: Introductionmentioning

confidence: 99%

Enhanced Colorimetric Signal for Accurate Signal Detection in Paper-Based Biosensors

Harpaz

Eltzov

et al. 2020

Diagnostics

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“…1,20 Even though lateral ow immunoassays 14,37,38 and other bioassays carried out in low-cost platforms 39,40 do not present such requirements, being faster and easier to use, pELISA testing tools overcome these testing tools because they present advantages such as a great multiplexing capacity and improved quantitative outputs, 20 and their low-cost also allows for broad-range screening tests in needy regions. These advantages are attainable only for tests with a high sensitivity (low number of false negative outputs), 41 as those demonstrated by our device aer the denition of its tolerance percentage.…”

Section: -36mentioning

confidence: 99%

“…19 However, the graphical analysis by itself is not capable of assessing the real performance of the new test. Moreover, a simple analytical curve and the determination of the conventional gures of merit of the new method (such as LOD and LOQ) will be dealing with negative outcomes, 20 which are not relevant for clinical applications.…”

Section: 814-16mentioning

confidence: 99%

“…20 We have opted for the later approach, in an attempt of eliminating the largest number of samples classied as false positive or false negative (Fig. 4B-F).…”

Section: -36mentioning

confidence: 99%

“…Thus, to determinate a safety margin to the outcomes of such low-cost testing devices, it is necessary to assess their performance indicators, 20 such as sensitivity (proportion of diseased people with a positive outcome), specicity (proportion of healthy people with a negative outcome) 21 and the accuracy (proportion of correct classications). 22 Moreover, a critical step for the establishment of a new diagnostic test is the determination of the cut-off value, which will dene the threshold value between healthy and diseased patients.…”

Section: 814-16mentioning

confidence: 99%

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Improved assessment of accuracy and performance indicators in paper-based ELISA

et al. 2017

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Paper-based devices are an excellent match for low-cost point-of-care testing (POCT) tools. Their userfriendliness, portability, and short time of analysis, coupled with ease of local manufacture make these devices the best option for inexpensive diagnostic testing tools. However, despite all their positive features, these low-cost diagnostic devices must present good performance indicators, such as sensitivity, specificity, and accuracy. We developed and validated a paper-based ELISA for toxoplasmosis diagnosis through the detection of Toxoplasma gondii immunoglobulin G (IgG) antibodies in 100 human serum samples. From among the different ways to define the cut-off value, we chose Youden's J index (cut-off ¼ 21.73 A.U.), which presented a higher sensitivity value. Our paper-based assay presented a sensitivity of 0.96, a specificity of 0.87, and a gray zone comprising 16 samples (AE15% of the cut-off value, with 3 false positive outputs). The accuracy of the test was estimated by using ROC curves (AUC ¼ 0.97). We also created a macro in Microsoft Excel® to estimate the accuracy of the test (m-Accuracy) based on a non-parametric method, which evidenced a value ¼ 0.88, which classifies our test as moderately to highly accurate. We also provide the m-Accuracy macro for download and the paperbased microplate designs for printing, in order to collaborate with the scientific community and facilitate further studies using this platform. The improvement of these diagnostic tools can bring this technology for those who need it, contributing to population health and well-being.

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Smart sampling as the “Spot‐on” Method for LC‐MS protein analysis from dried blood spots

Johannsen

Mrsa

Halvorsen

et al. 2023

J of Separation Science

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This perspective explores the feasibility of smart sampling with dried blood spots for the determination of proteins and peptides from human biomatrices using liquid chromatography coupled to mass spectrometry for clinical purposes. The focus is on innovative approaches to transform filter paper from a mere sample carrier to an active element in sample preparation, with the aim of reducing the need for extensive and intensive sample preparation in the conventional sense. Specifically, we discuss the use of modified cellulose to integrate sample preparation at an early stage of sample handling. The use of paper immobilized with either trypsin or monoclonal antibodies for protein digestion and affinity clean‐up is discussed as a potential benefit of starting sample preparation instantly at the moment of sampling to optimize time efficiency and enable faster analysis, diagnosis, and follow‐up of patients.

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Development and statistical assessment of a paper-based immunoassay for detection of tumor markers

Cited by 43 publications

References 44 publications

Enhanced Colorimetric Signal for Accurate Signal Detection in Paper-Based Biosensors

Enhanced Colorimetric Signal for Accurate Signal Detection in Paper-Based Biosensors

Improved assessment of accuracy and performance indicators in paper-based ELISA

Smart sampling as the “Spot‐on” Method for LC‐MS protein analysis from dried blood spots

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