Multiplexed Lateral Flow Assay with Segmented Nitrocellulose Membrane
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Solution Overview
Problem
Traditional lateral flow immunoassays are limited in their ability to detect multiple analytes simultaneously in a single sample, requiring sequential testing and being costly and time-consuming, which is not suitable for point-of-care or field use.
Innovation Solution
A multiplex lateral flow assay system with a nitrocellulose membrane having multiple assay test paths, where labeled detection reagents with low diffusion constants are sprayed onto the membrane, allowing for simultaneous detection of multiple analytes without physical barriers, enabling a compact, cost-effective, and easy-to-use device for detecting various analytes including infectious diseases and other biological markers.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional lateral flow immunoassays are used to detect multiple analytes, then sequential testing must be performed, but this increases testing time and cost
Solution Approach 1:
The membrane is divided into multiple independent assay test paths, each capable of detecting a different analyte simultaneously. This segmentation allows parallel processing of multiple analytes within a single test device, eliminating the need for sequential testing while maintaining the simplicity and low cost of traditional lateral flow assays
Solution Approach 2:
The lateral flow device is designed with multi-functionality to detect multiple analytes (e.g., SARS-CoV-2, influenza A, influenza B, RSV) simultaneously using a single sample application. Each test path contains specific capture reagents and labeled detection reagents tailored to its target analyte, enabling the device to perform multiple diagnostic functions in one test
2Adaptability or versatility
If multiple assay test paths are implemented on a single membrane, then simultaneous detection of multiple analytes is enabled, but device complexity increases
Solution Approach 1:
Each assay test path on the membrane possesses local quality with specific capture reagents and labeled detection reagents optimized for its target analyte. This localized specialization allows each path to function independently while maintaining overall system simplicity, as each path can be designed and validated separately before integration
Solution Approach 2:
The patent transitions from sequential one-dimensional testing to simultaneous multi-dimensional detection by arranging multiple assay test paths side-by-side on the membrane. This spatial dimensionality change allows multiple analytes to be detected in parallel without requiring multiple separate devices or complex sample handling procedures
3Measurement precision
If labeled detection reagents with low diffusion constants are used, then specificity of analyte detection is improved, but manufacturing precision requirements increase
Solution Approach 1:
The labeled detection reagents are pre-bound to specific locations on the membrane during manufacturing, creating fixed assay test paths with defined starting positions. This preliminary positioning ensures that when the sample flows through the membrane, the labeled detection reagents remain at precise locations, maintaining detection specificity without requiring high precision during the actual testing process
Solution Approach 2:
The patent utilizes labeled detection reagents with low diffusion constants as a key parameter change to maintain sharp, well-defined test lines. This parameter selection (low diffusion constant) ensures that even with variations in manufacturing precision, the reagents remain confined to their designated assay paths, preserving detection specificity while accommodating reasonable manufacturing tolerances
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables the simultaneous detection of multiple analytes in a single sample with high specificity and sensitivity, reducing costs and time, and is stable under various environmental conditions, making it suitable for point-of-care and field applications.
Implementation Method 1
When a liquid sample is applied to one end of the test membrane, the sample is drawn by capillary action along the longitudinal axis of the membrane strip
Implementation Method 2
Test line 16 contains analyte-specific molecules which are able to bind to and immobilize the analyte-conjugate complex, resulting in a visible colored line
Implementation Method 3
The conjugate consists of detection molecules specifically directed against the analyte of interest and indicator particles, such as colloidal gold or gold sol
Data Source
Figure 1~2B
Figure 3A~3B
Figure 3C~3D
AI summary
A device for performing a multiplex lateral flow immunoassay is provided in which a liquid sample, such as a biological sample, is simultaneously tested for the presence of multiple analytes of interest. Methods that employ the device in the simultaneous detection of multiple analytes of interest within a liquid test sample are also provided.