Biomolecule Analysis Kit with Low-Adsorption Reaction Wells
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Solution Overview
Problem
Existing biomolecule analysis methods, such as PCR and isothermal reactions, are either complicated or require long reaction times, making them impractical for rapid and quantitative analysis.
Innovation Solution
A biomolecule analysis kit and method utilizing an isothermal invader reaction in microspaces or wells with a surfactant-based adsorption inhibitor, such as Tween 20, to prevent reagent adsorption and enable rapid, quantitative analysis of DNA, RNA, and proteins.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR method is used for gene amplification and SNP detection, then detection accuracy is improved, but device complexity and operation complexity increase due to temperature control requirements
Solution Approach 1:
The invention changes the temperature parameter from variable (PCR method requiring heating and cooling cycles) to constant (isothermal reaction at 60-65°C), thereby simplifying the device structure while maintaining SNP detection accuracy through the invader reaction mechanism
Solution Approach 2:
The invention replaces the mechanical thermal cycling system with a chemical isothermal reaction system, eliminating the need for complex temperature control mechanisms while achieving the same analytical goal through the invader reaction's constant-temperature amplification
2Measurement precision
If two-stage operation (gene amplification + SNP detection) is performed, then SNP detection accuracy is improved, but operation complexity and time consumption increase
Solution Approach 1:
The invention merges the gene amplification function and SNP detection function into a single isothermal invader reaction process, where the reaction simultaneously amplifies the target sequence and generates detectable signals that directly indicate SNP presence, eliminating the need for separate amplification and detection stages
Solution Approach 2:
The invader reaction system performs multiple functions simultaneously: it acts as both the gene amplification mechanism and the SNP detection mechanism, with the reaction products serving dual purposes as both amplified DNA and detection signals, thereby simplifying the overall operational procedure
3Device complexity
If isothermal reaction without PCR amplification is used, then device complexity is reduced, but signal amplification speed decreases and reaction time increases
Solution Approach 1:
The invention optimizes the reaction temperature parameter to 60-65°C, which is higher than typical isothermal reactions, thereby accelerating the enzymatic reaction rate and signal amplification speed while maintaining the simplicity of isothermal operation without PCR's thermal cycling
Solution Approach 2:
The invention introduces an intermediary signal amplification mechanism where the invader reaction produces detectable signals (such as fluorescence or colorimetric changes) that amplify the detection response, compensating for the lack of exponential PCR amplification while keeping the device structure simple
4Quantity of substance
If reagent adsorption onto well surfaces occurs, then reagent concentration is reduced, but reaction accuracy and reproducibility deteriorate
Solution Approach 1:
The invention introduces a surfactant as an intermediary substance that adsorbs onto the well surfaces, forming a protective layer that prevents reagent components from adhering to the surface, thereby maintaining reagent concentration and ensuring consistent reaction results across multiple replicates
Solution Approach 2:
The surfactant creates a standardized surface environment by uniformly coating all well surfaces, ensuring that each reaction well has identical surface properties, which eliminates variability caused by surface adsorption and improves reaction reproducibility
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
The method allows for rapid and quantitative analysis of biomolecules with improved reactivity, reducing reaction time and reagent consumption while maintaining high reproducibility and accuracy.
Implementation Method 1
a biomolecule analysis kit and method utilizing an isothermal invader reaction in microspaces or wells with a surfactant-based adsorption inhibitor, such as Tween 20, to prevent reagent adsorption
Data Source
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Figure 3
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AI summary
A biomolecule analysis kit includes a reaction container configured to perform an enzymatic reaction, the reaction container including a base portion which has a container-shaped portion and a low-adsorption structural portion which is provided on at least the inner surface of the container-shaped portion, the low-adsorption structural portion having an adsorption rate lower than the base portion at which at least one of a sample which becomes a target of analysis in the enzymatic reaction and a reagent for the enzymatic reaction is adsorbed thereonto, wherein a signal resulting from the enzymatic reaction is configured to be detected when the enzymatic reaction is performed in the reaction container.