Ionic Liquid PCR Inhibitor Suppression for Stool DNA
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Nucleic acid amplification from biological materials is hindered by the presence of inhibitors such as heme, bilirubin, and complex polysaccharides, particularly in stool samples, where existing removal methods are ineffective and can result in loss of target cells or DNA.
Innovation Solution
The use of ionic liquids in the nucleic acid amplification process, which suppresses the activity of nucleic acid amplification inhibitors, thereby improving reaction efficiency and sensitivity by enhancing the polymerase chain reaction (PCR) efficiency and reducing the presence of inhibitors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Object-affected harmful factors
If conventional removal methods are used to eliminate nucleic acid amplification inhibitors from biological materials, then inhibitor removal is attempted, but target cells or DNA are lost and removal effectiveness is insufficient
Solution Approach 1:
Ionic liquids serve as intermediary substances that directly suppress the activity of nucleic acid amplification inhibitors (heme, bilirubin, bile salts, polysaccharides) without requiring complex pre-treatment steps. The ionic liquid molecules interact with the inhibitors to neutralize their harmful effects, allowing PCR to proceed efficiently without losing target cells or DNA.
Solution Approach 2:
The invention changes the chemical environment parameters by introducing ionic liquids with specific properties (charge, polarity, viscosity) into the PCR reaction system. This parameter change allows the reaction to tolerate the presence of inhibitors that would normally prevent amplification, effectively suppressing inhibitor activity without removing the inhibitors themselves.
2Object-affected harmful factors
If pre-treatment methods are used to remove inhibitors before PCR preparation, then inhibitor removal is attempted, but the process becomes complicated and target cells may be lost
Solution Approach 1:
The ionic liquid is added directly to the PCR master mix before the amplification reaction begins, performing the inhibitor suppression action in advance. This preliminary action eliminates the need for separate, complicated pre-treatment steps while ensuring inhibitors are suppressed before they can interfere with the PCR process.
Solution Approach 2:
The ionic liquid acts as a mediator substance that bridges the incompatible elements (inhibitors and PCR reaction). By introducing this intermediary, the system can tolerate inhibitor presence without requiring complex separation or removal procedures, significantly simplifying the overall process.
3Loss of substance
If post-treatment methods are used to add inhibitor removal substances to PCR master mix, then target cell loss is avoided, but inhibitor removal effectiveness remains insufficient
Solution Approach 1:
The invention uses ionic liquids with specific physicochemical parameters (ionic character, polarity, molecular size) that enable effective inhibitor suppression. By changing the chemical environment through ionic liquid addition, the system achieves superior inhibitor removal effectiveness while maintaining target cell integrity, overcoming the limitations of conventional post-treatment substances.
Solution Approach 2:
The PCR master mix is transformed into a composite system by incorporating ionic liquids alongside traditional PCR reagents. This composite formulation combines the functions of DNA polymerase, buffers, dNTPs, primers, and ionic liquids, creating a synergistic system that achieves both target cell preservation and effective inhibitor suppression.
4Ease of operation
If conventional PCR is performed in the presence of inhibitors, then the process is simple, but amplification efficiency and sensitivity are reduced
Solution Approach 1:
By introducing ionic liquids that change the chemical parameters of the reaction environment (ionic strength, polarity, viscosity), the system maintains operational simplicity while dramatically improving amplification efficiency and sensitivity. The ionic liquid creates an optimized chemical environment that enhances PCR performance without adding operational complexity.
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 effectively amplifies nucleic acids in the presence of inhibitors, improving PCR efficiency and sensitivity, allowing for rapid and precise detection of target microorganisms or genes without significant loss of target cells or DNA.
Implementation Method 1
amplifying the nucleic acid amplification in the presence of an ionic liquid
Implementation Method 2
suppresses the activity of the nucleic acid amplification inhibitor
Data Source
AI summary
A method for amplifying a nucleic acid amplification in the presence of an ionic liquid that suppresses an inhibitor of nucleic acid amplification, particularly when in a biological material, and a composition useful for performing the method.