Universal Membrane Nucleic Acid Detection Kit
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Solution Overview
Problem
Conventional molecular diagnosis techniques, such as PCR, face challenges in clinical laboratories due to the complexity and cost of equipment and reagents, particularly in performing multiplex PCR and confirming amplified DNA, which limits their use in general clinical settings.
Innovation Solution
A method and kit for detecting target nucleic acids using a primer with a binding region complementary to the target nucleic acid, a nucleic acid oligomer non-complementary to it, and a labeled primer, reacting the amplified product with a membrane probe for universal identification, allowing for simultaneous detection of multiple targets using a single membrane.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional PCR techniques are used with electrophoresis for DNA confirmation, then DNA amplification can be achieved, but the procedure becomes complicated and unsuitable for clinical laboratories
Solution Approach 1:
The invention extracts and eliminates the electrophoresis step from the conventional PCR workflow. By incorporating a label directly onto the primer and using a membrane-based detection system with a probe, the method removes the need for gel electrophoresis, agarose preparation, and DNA staining, thereby simplifying the procedure while maintaining reliable DNA amplification and detection
Solution Approach 2:
The invention introduces a labeled primer and a membrane-bound probe as intermediary elements. The label on the primer serves as a marker that can be detected without electrophoresis, and the probe on the membrane acts as an intermediary that specifically binds to the amplified DNA, enabling direct detection and eliminating the need for complex electrophoresis equipment
2Device complexity
If real-time PCR techniques are used, then electrophoresis is not required, but expensive instruments and fluorescent reagents are needed
Solution Approach 1:
The invention employs a cost-effective labeling approach using labels on primers that can be detected on a simple membrane, replacing expensive fluorescent reagents and instruments. The method uses affordable materials such as labeled primers and membrane probes, making the detection process economically viable for routine clinical use without requiring expensive real-time PCR equipment
Solution Approach 2:
The invention replaces the complex optical detection system of real-time PCR (fluorescence detection requiring expensive instruments) with a simpler membrane-based detection system. The label on the primer is detected through binding with the membrane probe, eliminating the need for expensive fluorescent detection equipment while achieving similar detection goals
3Measurement precision
If real-time PCR with limited fluorescence wavelengths is used, then detection can be performed, but multiplex PCR for five or more samples is difficult
Solution Approach 1:
The invention creates a universal membrane that can detect multiple different targets simultaneously. By using a single membrane with probes that bind to different labeled primers, the system achieves multiplex detection capability without being limited by fluorescence wavelengths. The membrane serves multiple functions by detecting various targets through the universal binding mechanism of the probe-label interaction
Solution Approach 2:
The invention changes the detection parameter from fluorescence wavelength (which has limited multiplexing capability) to label type or position on the membrane, which can accommodate multiple targets. By using different labels or different probe binding positions on the membrane, the system can distinguish and detect five or more different targets simultaneously, greatly enhancing multiplex capability
4Device complexity
If nucleic acid lateral flow assay is used with membrane instead of gel electrophoresis, then procedure is simplified, but probe sequence must be specifically designed for each PCR product
Solution Approach 1:
The invention creates a universal membrane that can be used for detecting different PCR products without requiring custom probe design for each target. The membrane probe is designed to bind to a common region or structure present in all targets, or the system uses a panel of standardized probes that can detect multiple targets. This universality simplifies manufacturing and eliminates the need for complex custom probe design for each PCR product while maintaining procedure simplicity
Solution Approach 2:
The invention performs preliminary design of universal probes or label systems that can detect multiple targets. By pre-designing the membrane probe to recognize common features or by establishing a standardized set of probes for common target regions, the system eliminates the need for custom probe design for each new application, thereby simplifying both manufacturing and usage
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 simplifies and enhances the accuracy of nucleic acid analysis, reducing production costs and enabling simultaneous diagnosis of multiple targets with a universal membrane, making it more suitable for clinical use.
Implementation Method 1
amplifying the target nucleic acid by reacting a sample comprising the target nucleic acid with a primer comprising (i) a binding region complementary to the target nucleic acid
Implementation Method 2
reacting a product amplified in step (a) with a membrane having immobilized thereon a probe that binds complementarily to the nucleic acid oligomer
Data Source
AI summary
The present disclosure relates to a method and a kit for analyzing a nucleic acid amplification product, and more particularly to a method and kit of analyzing a nucleic acid amplification product using a labeled primer comprising (i) a binding region complementary to the target nucleic acid, (ii) a nucleic acid oligomer containing nucleotides non-complementary to the target nucleic acid, and (iii) a label, and using a membrane having immobilized thereon a probe that binds complementarily to the nucleic acid oligomer.