Lateral Flow Protein Detection via Nucleic Acid Amplification
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Solution Overview
Problem
Detecting target proteins at low concentrations in biological samples poses challenges due to sensitivity limitations and laborious concentration procedures, especially in point-of-care applications where sample volumes are limited.
Innovation Solution
The use of a target-linked template nucleic acid generation technique, such as proximity assays, to amplify nucleic acids for detection on lateral flow substrates, enabling high sensitivity detection of proteins at low concentrations within small sample volumes through a fluidic device that automates fluidic flow and processing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional lateral flow substrates are used to bind target proteins, then the detection method is simple and rapid, but the sensitivity is insufficient to achieve accurate diagnostic indications
Solution Approach 1:
The patent introduces nucleic acid amplification as an intermediary step between protein binding and detection. The lateral flow substrate first binds the target protein, then a nucleic acid probe binds to the protein and undergoes amplification, which serves as a mediator to amplify the signal and achieve high sensitivity detection while maintaining the simplicity of the lateral flow format.
Solution Approach 2:
The patent changes the detection parameter from direct protein binding signal to nucleic acid amplification signal. By converting the protein binding event into a nucleic acid-based detection process with amplification capability, the system achieves enhanced sensitivity while preserving the rapid and simple lateral flow substrate methodology.
2Measurement precision
If sample concentration procedures are performed to achieve suitable concentration for detection, then detection sensitivity is improved, but the procedures become laborious and time-consuming
Solution Approach 1:
The patent incorporates sample concentration functionality directly into the lateral flow device structure, performing the concentration action preliminarily within the device itself rather than requiring separate external concentration steps. This integrated approach eliminates laborious manual concentration procedures while maintaining the ability to detect low-concentration proteins.
Solution Approach 2:
The patent merges the sample concentration function with the detection function in a single integrated lateral flow device. The device simultaneously performs sample concentration, protein binding, and signal detection in one unified system, eliminating the need for separate concentration procedures and reducing operational complexity.
3Adaptability or versatility
If conventional detection methods are used with limited sample volumes, then point-of-care applicability is improved, but accurate protein detection becomes difficult
Solution Approach 1:
The patent uses nucleic acid amplification to create multiple copies of the detection signal from a single protein binding event. This copying mechanism allows the system to amplify the signal from limited sample volumes, enabling accurate protein detection in point-of-care applications where only small sample amounts are available.
Solution Approach 2:
The patent transitions from direct protein detection in the original sample dimension to nucleic acid-based detection in an amplified dimension. By converting the protein binding signal into nucleic acid copies that can be amplified, the system overcomes the limitation of small sample volumes while maintaining point-of-care applicability.
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
This approach allows for accurate detection of proteins at concentrations as low as 10 pg/mL and reduces sample and reagent handling, enabling rapid and reliable point-of-care diagnostics with minimal sample volume requirements.
Implementation Method 1
use of lateral flow substrates and technologies to bind target proteins
Implementation Method 2
target-linked template nucleic acid generation technique, such as proximity assays
Data Source
AI summary
Methods, systems, and devices for analyzing a sample for the presence of a target protein are provided. A sample can be processed, the processing generating an amplification product of a template nucleic acid formed in response to a target protein being present in the sample. The amplification product can be contacted with a detection probe to generate a second product comprising a labeled nucleic acid. A lateral flow substrate can be contacted with the second product, the lateral flow substrate comprising a capture moiety configured to bind the labeled nucleic acid. The methods can be carried out in one or more chambers of a single device or multiple devices. Detection probe bound to amplicons of the template nucleic acid captured on the lateral flow substrate can be read visually or using a sensor.