Reaction Card Micropump Sampling for Nucleic Acid Detection
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Solution Overview
Problem
Nucleic acid amplification methods are prone to false positive results due to sample contamination and require substantial sampling volumes, limiting their sensitivity and efficiency in detection and quantification.
Innovation Solution
A system comprising a reaction card with channel networks, valves, and micropumps, along with a reaction vessel assembly, allows for real-time, quantitative PCR by sampling and analyzing amplification products in a controlled manner, minimizing contamination and sampling volume through a reagent tube assembly and tubular members for fluid communication.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If nucleic acid amplification methods are used to improve detection sensitivity, then detection sensitivity is improved, but sample contamination risk increases leading to false positive results
Solution Approach 1:
The system divides the amplification reaction into multiple discrete sampling points throughout the reaction cycles. Aliquots are taken at different time points (e.g., after 10, 20, 30 cycles) rather than sampling the entire reaction mixture at once. This segmentation allows detection of amplification products while minimizing the volume subject to contamination and reducing false positives.
Solution Approach 2:
The system performs preliminary sampling and analysis of amplification products during the amplification reaction itself, before the reaction is complete. By taking aliquots at intermediate cycles and analyzing them in real-time, the system can detect target nucleic acids early, reducing the need for large final sampling volumes and minimizing contamination exposure.
2Reliability
If large sampling volumes are used to ensure adequate detection, then detection reliability is improved, but the complexity of the sampling system increases
Solution Approach 1:
The system extracts only the necessary aliquots of amplification product at specific time points during the reaction, rather than requiring analysis of the entire reaction mixture. This extraction approach maintains detection reliability by obtaining sufficient sample material while simplifying the overall sampling system design and reducing the volume that requires handling and storage.
Solution Approach 2:
The system takes partial samples (aliquots) of the amplification product rather than sampling the complete reaction mixture. By taking multiple small aliquots at different time points rather than one large sample, the system achieves reliable detection with reduced sampling volume and simplified system requirements.
3Measurement precision
If multiple sampling points are taken during amplification cycles to improve quantification accuracy, then quantification accuracy is improved, but the time required for analysis increases
Solution Approach 1:
The system performs sampling and analysis continuously during the amplification reaction cycles without stopping the reaction. Multiple aliquots are taken at different cycle points (e.g., cycles 10, 20, 30) while the amplification continues, allowing quantification accuracy to improve through multiple data points while minimizing additional analysis time by overlapping sampling with ongoing amplification.
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 system enhances the sensitivity of nucleic acid detection while reducing contamination risks and sampling volume, enabling more precise and efficient analysis of nucleic acids during amplification reactions.
Implementation Method 1
the micropump is activated to pump a first aliquot of reaction product from the reaction vessel into the tubular member, through the channel network, and into the collection well
Implementation Method 2
the valve is opened to atmosphere, and the micropump is activated to pump a first aliquot of reaction product from the reaction vessel
Data Source
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AI summary
The invention provides systems and methods for processing samples. In a method, a reaction card is provided that has a channel network, a valve, and a micropump, all disposed within the card. The reaction card also has a collection well disposed on a surface of the card and a tubular member extending out from the card. A reaction vessel is provided and affixed to the reaction card such that the tubular member is inserted into the reaction vessel. Amplification reaction reagents and a sample are delivered into the reaction vessel, and an amplification reaction is initiated within the reaction vessel, resulting in an amplification product being disposed within the reaction vessel. The valve is opened to atmosphere, and the first micropump is activated to pump an aliquot of reaction product from the reaction vessel into the tubular member, through the channel network, and into the collection well.