Isothermal Nucleic Acid Detection with Fluid-Tight Chamber Assembly
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Solution Overview
Problem
Current nucleic acid amplification technologies, such as PCR, require thermal cycling and sophisticated equipment, whereas isothermal methods like RPA offer advantages but lack efficient sample preparation and contamination-free handling for point-of-care applications.
Innovation Solution
A set of containers with distinct chambers for lysis and amplification, using recombinase polymerase amplification (RPA) enzymes and a fluorescence detection system, allowing for isothermal nucleic acid amplification and minimizing contamination through a fluid-tight assembly and snap-fit connection, enabling easy handling and disposal.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Device complexity
If isothermal nucleic acid amplification (RPA) is used, then equipment complexity is reduced and room temperature operation is achieved, but contamination risk increases and sample preparation efficiency is insufficient
Solution Approach 1:
The system is divided into separate functional modules: a lysis container for sample preparation and a test container for amplification and detection. This segmentation allows each module to be optimized independently, with the lysis container providing robust sample processing and the test container providing contamination-free amplification in a controlled environment.
Solution Approach 2:
A fluid transfer interface acts as an intermediary between the lysis container and test container. This interface includes a valve mechanism that controls fluid transfer, ensuring that lysed sample is transferred to the test container without contamination while maintaining the simplicity of the overall system.
2Reliability
If separate containers for lysis and amplification are used, then contamination is minimized, but handling complexity increases
Solution Approach 1:
The lysis container and test container are designed to be combined through a snap-fit connection mechanism. When connected, they form an integrated assembly where the lysis container's fluid transfer interface automatically interfaces with the test container's receiving structure, simplifying the transfer process while maintaining contamination-free separation during handling.
Solution Approach 2:
The fluid transfer interface includes an automatically actuating valve that opens when the containers are connected and closes when disconnected. This self-regulating mechanism eliminates the need for manual valve control, reducing handling complexity while ensuring contamination-free transfer.
3Device complexity
If manual fluid transfer is used between containers, then equipment simplicity is maintained, but transfer precision and contamination control are insufficient
Solution Approach 1:
The fluid transfer interface utilizes pressure differential and fluid pressure to control the transfer process. When the containers are connected, pressure differential automatically opens the valve to transfer lysed sample to the test container, and pressure equalization automatically closes the valve, providing precise control without complex mechanical components.
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
Facilitates rapid, contamination-free nucleic acid detection at room temperature, reducing equipment complexity and improving sample reliability, especially suitable for point-of-care testing and simultaneous detection of DNA and RNA without separate cDNA production steps.
Implementation Method 1
the mixture comprises a recombinase, a single-stranded DNA-binding protein (SSB) and strand-displacing polymerase that causes a recombinase polymerase amplification (RPA)
Implementation Method 2
After the light source at specific wavelength illuminates on the targeted nucleic acids, the DNA-binding dyes or fluorescein- binding probes of the nucleic acids will react and enable fluorescent signals to be emitted
Implementation Method 3
The lysis container may contain a liquid lysing fluid that causes lysing of the cells in a sample to thus release the nucleic acids (DNA or RNA)
Data Source
Figure 1
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Figure 3A~3B
AI summary
The invention relates to a system is provided that comprises a first container, a second container and a fluorescence detection device. The first container comprises a first set of chemicals and/or agents and is closed prior to use. The second container comprises a second set of chemicals and/or agents that are at least in part distinct from the chemicals and/or agents of the first set. The first container comprises a lid, that can be opened when the first container and the second container are combined to form a single, fluid tight assembly, in order to allow the contents of the first container to enter the second container.