Magnetic Microparticle Nucleic Acid Extraction for Integrated Amplification
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Solution Overview
Problem
Existing methods for detecting nucleic acids in samples have limited sensitivity, requiring high concentrations and often necessitate separate extraction and amplification steps, which can be inefficient and impractical.
Innovation Solution
A method and device utilizing magnetic microparticles functionalized with extraction nucleic acids and primers, allowing for hybridization, separation, and localized heating to enhance nucleic acid extraction and amplification efficiency.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If magnetic microparticles are used for extraction and amplification, then detection sensitivity is improved, but device complexity increases
Solution Approach 1:
The patent combines extraction and amplification functions into a single integrated system using magnetic microparticles. The microparticles serve multiple purposes: they extract target nucleic acids through hybridization, concentrate them via magnetic separation, and facilitate amplification when placed in the heating element, eliminating the need for separate extraction and amplification steps
Solution Approach 2:
The magnetic microparticles are designed with multi-functionality, serving as both extraction carriers and amplification substrates. They are functionalized with nucleic acid binding capabilities for extraction and can be directly utilized in PCR amplification when positioned in the heating element, reducing the number of required components
2Reliability
If separate extraction and amplification steps are performed, then detection reliability is improved, but processing time increases
Solution Approach 1:
The patent merges extraction and amplification into a single continuous process. Magnetic microparticles extract target nucleic acids from the sample matrix, and without requiring separate purification steps, are directly transferred to and used in the amplification reaction within the same reaction vessel, significantly reducing processing time while maintaining reliability
Solution Approach 2:
The magnetic microparticles perform preliminary extraction and concentration of target nucleic acids within the reaction vessel before amplification begins. This preliminary action prepares the sample in-situ, eliminating the need for separate extraction procedures and reducing overall processing time
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
Enables efficient extraction and amplification of nucleic acids with improved detection limits by facilitating hybridization and separation using magnetic forces, while minimizing interference from the sample matrix.
Implementation Method 1
providing a magnetic field in the reaction vessel such that at least a portion of the magnetic microparticles bound to the target nucleic acid attach to an extraction element arranged in and/or on the reaction vessel
Implementation Method 2
locally heating the reaction solution to a denaturation temperature by means of the local heating element in the region in which the magnetic microparticles are attached to the local heating element
Implementation Method 3
hybridizing at least a portion of the target nucleic acid with one of the extraction nucleic acids and binding the target nucleic acid via the extraction nucleic acid to one of the magnetic microparticles
Data Source
Figure 1~2
Figure 3A~3D
Figure 4A~4D
AI summary
The invention relates to a method for extracting a target nucleic acid (12) from a sample liquid (30). The method comprises the provision of a sample liquid (30) with the target nucleic acid (12) in a reaction vessel (22) and the provision of magnetic microparticles (10) in the sample liquid (30), which are in each case functionalized with at least one extraction nucleic acid (16), wherein the extraction nucleic acids (16) are at least partially complementary to the target nucleic acid (12). The method further comprises a hybridization of at least a part of the target nucleic acid (12) with one of the extraction nucleic acids (16) and binding the target nucleic acid (12) via the extraction nucleic acid (16) to one of the magnetic microparticles (10). Moreover, the method comprises the provision of a magnetic field in the reaction vessel (22) such that at least a part of the magnetic microparticles (10) bound to the target nucleic acid (12) is sedimented on a vessel wall (22a, 22b, 22c) of the reaction vessel (22). Furthermore, devices and methods for amplifying a target nucleic acid (12), the use of magnetic microparticles (10) for extracting a target nucleic acid (12) and a magnetic microparticle are provided.