Portable Paper-Based Immunoassay Combined with Smartphone Application for Colorimetric and Quantitative Detection of Dengue NS1 Antigen
January 26th, 2024
Addressing urgent dengue diagnostic needs, here we introduce a smartphone app-integrated Dengue NS1 Paper-based Analytical Device (DEN-NS1-PAD) for quantifying Dengue NS1 antigen concentration in clinical serum/blood samples. This innovation enhances dengue management by aiding clinical decision-making in various healthcare settings, even resource-limited ones.
The scope of the research is detecting dengue using a specific biomarker, NS1 with a paper-based analytical device. Our research focus on developing a simple, cheap, sensitive, and specific device to detect dengue. We also developed smartphone application to improve sensitivity and measurement of NS1 in real sample.
Our sensitivity is developing a method to fabricate microfluidic method based on our PEC Using formula This innovative techniques, aims to embody commercialization of various PEC designs, offering scalable solutions for applications. Our research combines PEC with smartphone application, which is very promising for dengue NS1 measurement. The mobile application significantly enhanced sensitivity and efficiency in qualifying NS1 in serum samples, compared to observation with the naked eye.
This protocol is simple, cheap, and reproducible to produce PEC to detect dengue NS1. The PEC is sensitive and specific for detecting dengue NS1, and will increase the sensitivities after combining with the smartphone. Our goal is to improve dengue NS1 test performance with the blood samples, evaluating the cross-infections cell types, and incubating it into the paper-based devices for multiple diseases.
Addressing technology culture trend is crucial for successful commercialization in bring all the disease screening and medical care. To begin, design a paper analytical device with 18 wax patterns on a computer. Print this design onto a cellulose paper using a wax printer.
Melt the wax printed paper in a laboratory oven for 75 seconds at 150 degrees Celsius. After melting, store the paper in a silica box until further use. Next pipette 0.5 microliters of 0.025%polylysine to both the test and control areas of the device.
Then incubate the device in the oven at 65 degrees Celsius for five minutes. Apply 0.5 microliters of the respective antibody solution to the control and test areas. Allow the drops to dry at room temperature in a silica gel box for 30 minutes.
Next, apply the blocking buffer to the sample conjugate and detection areas. Let these drops dry at room temperature in a silica gel box for 30 minutes. Combine 10 microliters of anti NS1 one antibody in PBS with one milliliter of 40 nano molar gold nanoparticle colloid.
Then add 0.1 milliliter of 0.1 molar borate buffer. Rotate the mixture at 50 RPM for 60 minutes. Pipette 0.1 milliliter of BSA in BBS into the mixture.
Rotate again before incubation. Centrifuge the solution at 20, 187 G at four degrees Celsius for 30 minutes. Then carefully supernatant from the precipitate gold nanoparticle antibody solution.
Resuspend the precipitate in 500 microliters of BBS. Next, sonicate the solution to disperse the nanoparticles. After centrifuging, add 50 microliters of the conjugate buffer to the suspension.
Then pipette two microliters of gold nanoparticle antibody complex solution to the conjugate area. To assemble the paper-based immunoassay carefully remove the protective film from the reverse side of the adhesive plastic backing card to expose the adhesive. Align the treated cellulose paper with the adhesive plastic backing card and press.
Apply a plastic film over the paper and press the film and paper together to create a coated surface. A single channel immunoassay device was designed and fabricated the sample solution wicked through the channel and interacted with the components at key points resulting in a visible diagnostic result. To begin, transfer 300 microliters of peripheral blood from 30 patients on the first day of hospitalization into purple top EDTA tubes.
Centrifuge the blood at 2884 G at four degrees Celsius for 20 minutes. Transfer the plasma into a clean polypropylene tube. Next, apply 20 microliters of plasma onto the sample area of the immunoassay device.
Now pipette 30 microliters of the wash buffer to the sample area. After allowing the sample to completely waken to the device, capture the images of the device with a smartphone. Analyze the test and control areas using image J and the custom mobile application to quantify the intensity of the assay in the mobile application.
Launch the developed mobile application on the smartphone, select use camera or upload from gallery to choose or upload the data source. Now navigate to the analytics section and press analyze. The application will now process the data and display the results.
The paper-based immunoassay device was able to rapidly diagnose dengue, similar to a commercially available diagnostic test.
Overview
This study presents a smartphone app-integrated Dengue NS1 Paper-based Analytical Device (DEN-NS1-PAD) designed for quantifying Dengue NS1 antigen concentration in clinical serum samples. The device aims to enhance dengue management by improving clinical decision-making, particularly in resource-limited healthcare settings.
Key Study Components
Area of Science
- Neuroscience
- Biotechnology
- Diagnostic Devices
Background
- Dengue fever is a significant global health concern.
- Current diagnostic methods may be inadequate in resource-limited settings.
- NS1 antigen is a critical biomarker for dengue detection.
- There is a need for simple, cost-effective diagnostic solutions.
Purpose of Study
- To develop a sensitive and specific device for dengue detection.
- To integrate a smartphone application to enhance measurement accuracy.
- To provide scalable solutions for dengue diagnostics.
Methods Used
- Development of a paper-based analytical device (PAD).
- Fabrication of microfluidic methods based on PEC.
- Integration of a smartphone application for enhanced sensitivity.
- Testing the device with real serum samples.
Main Results
- The DEN-NS1-PAD demonstrated improved sensitivity in detecting NS1.
- The smartphone application significantly enhanced measurement efficiency.
- Results showed better performance compared to traditional observation methods.
- The device is suitable for use in various healthcare settings.
Conclusions
- The DEN-NS1-PAD is a promising tool for dengue diagnostics.
- Integration with mobile technology offers significant advantages.
- This innovation can improve clinical decision-making in dengue management.
Frequently Asked Questions
Executive Industry Relevance
Quantitative detection of dengue NS1 antigen using a portable paper-based immunoassay with smartphone integration addresses a critical need for sensitive, scalable diagnostics in both centralized and resource-limited settings. This approach enhances predictive confidence in early infection detection and supports rapid triage decisions, directly impacting portfolio strategies for infectious disease diagnostics. The integration of digital quantification elevates assay reproducibility and enables broader enterprise deployment across diverse healthcare environments.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables direct quantification of NS1 antigen, supporting robust biomarker validation.
- Facilitates biological de-risking by providing sensitive, specific detection in real clinical samples.
- Improves predictive confidence for downstream diagnostic and therapeutic development.
Screening & Assay Development
- Delivers standardized, reproducible colorimetric outputs suitable for high-throughput screening.
- Supports assay scalability and platform reuse through simple fabrication and digital readout.
- Enables reliable evaluation of compound or antibody performance in diagnostic workflows.
Translational & Preclinical Research
- Aligns with translational biomarker strategies by quantifying NS1 in clinically relevant matrices.
- Provides continuity from discovery to preclinical validation for infectious disease diagnostics.
- Reduces risk in advancing diagnostic candidates by enabling rapid, field-deployable testing.
Pipeline & Workflow Integration
This portable immunoassay system fits from early discovery through lead identification and preclinical validation for infectious disease diagnostics.
- Discovery Biology: Supports hypothesis testing and pathway clarification via quantitative NS1 measurement.
- Screening: Offers reproducible, digital assay outputs for comparative analysis across samples.
- Analytics: Provides quantitative colorimetric data for robust statistical evaluation of diagnostic performance.
- Translational Research: Bridges laboratory and field settings, supporting biomarker alignment and clinical relevance.
- Enterprise Reuse: Establishes a scalable, adaptable platform for multi-disease screening and rapid deployment.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in NS1 detection.
- Operational Value: Enhances standardization, reproducibility, and scalability of diagnostic assays.
- Strategic Value: Enables faster go/no-go decisions and improves capital efficiency in diagnostic development.
- Portfolio Impact: Supports risk-adjusted prioritization and advancement of diagnostic candidates.
Implementation Considerations
- Requires expertise in immunoassay development and digital image analysis.
- Needs access to basic laboratory instrumentation and smartphone-based analytics infrastructure.
- Demands cross-team standardization for assay fabrication and data interpretation.
- Adaptable to various sample types and potentially other disease biomarkers.
- Performance may vary with sample matrix and environmental conditions, requiring validation.
