Porous Structure Primer Fixation Multiplex PCR
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Solution Overview
Problem
Current multiplex real-time PCR methods face challenges in accurately and rapidly analyzing multiple nucleic acids simultaneously due to interference between targets, making it difficult for accurate disease diagnosis in fields like point-of-care technology.
Innovation Solution
A porous structure with primers fixed inside its pores, made from a hydrophilic polymer with 60-80% porosity, allowing for real-time amplification and detection of various nucleic acids by varying the primers included in the structure, enhancing reactivity and enabling simultaneous analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If multiplex real-time PCR is conducted using probe color and melting point methods, then various biomarkers can be confirmed in a single chamber, but measurement accuracy deteriorates due to interference between targets as the number of targets increases
Solution Approach 1:
The invention divides the detection system into separate porous structures, each containing specific primers for different target nucleic acids. This segmentation allows multiple targets to be analyzed simultaneously in a single chamber while preventing interference between targets, as each porous structure independently captures and amplifies its specific target.
Solution Approach 2:
The porous structures act as intermediaries between the target nucleic acids and the detection system. These structures with controlled porosity (60-80 vol%) and specific primer compositions facilitate selective binding and amplification of target sequences while blocking interference from non-target substances, thereby maintaining measurement accuracy in multiplex applications.
2Ease of manufacture
If endpoint PCR is used for nucleic acid amplification, then the method is simple and commonly used, but real-time detection and quantification of nucleic acid cannot be performed
Solution Approach 1:
The invention merges the amplification and detection functions into a single integrated system. The porous structures containing primers enable real-time monitoring of nucleic acid amplification throughout the PCR process, eliminating the need for separate endpoint analysis steps while maintaining the simplicity of the PCR methodology.
Solution Approach 2:
The porous structures enable continuous monitoring of the PCR reaction in real-time throughout the amplification process. This continuous detection capability allows for real-time quantification of nucleic acids during the exponential phase of amplification, eliminating the time loss associated with post-reaction analysis while maintaining procedural simplicity.
3Productivity
If the porosity of the porous structure is increased to maximize reactivity and utilize interior space, then the structure can better contain primers and enable real-time amplification, but the structural stability may be compromised
Solution Approach 1:
The invention optimizes the porosity parameter of the porous structures to a specific range (60-80 vol%) that balances reactivity and structural stability. This parameter optimization ensures sufficient interior space for primer containment and real-time amplification while maintaining adequate structural integrity for practical application.
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 porous structure enables real-time detection and quantification of multiple nucleic acids, improving the accuracy and speed of disease diagnosis, particularly in point-of-care settings by maximizing reactivity and utilizing the interior space for amplification.
Implementation Method 1
a porous structure including pores and primers in one or more directions between a forward primer and a reverse primer of a target nucleic acid as a primer of polymerase chain reaction (PCR) fixed to the interior of the pores
Implementation Method 2
the porous structure is a cured pre-polymer which is a hydrophilic polymer, wherein a porosity of the porous structure is from 60 vol% to 80 vol% with respect to a total volume of the porous structure
Data Source
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AI summary
A porous structure according to the present invention has a polymerase chain reaction (PCT) primer inside pores thereof, and hence, even an inner portion thereof can be used unlike general structures of which only surfaces are used for amplification and detection, thereby maximizing reactivity. In addition, the differentiating of the kinds of primers contained in respective structures leads to detection of several kinds of target nucleic acids at the same and real-time analysis thereof at the same time, and thus is useful for multiplex real-time PCR.