Nucleic Acid Extraction via High-Pressure Heating
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Solution Overview
Problem
Conventional nucleic acid extraction methods from biological samples, particularly from paraffin-embedded tissues, are time-consuming, inefficient, and require multiple steps and specialized equipment, limiting the speed and cost-effectiveness of gene detection processes.
Innovation Solution
A method involving high-pressure heating of biological samples, optionally with a dissolution medium, to release nucleic acids into a liquid phase, simplifying the extraction process and allowing direct use of the resulting solution for detection methods like PCR, without the need for extensive dewaxing or lysing steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional chemical methods (enzyme digestion, organic solvent extraction, ethanol precipitation) are used to extract nucleic acids from biological samples, then nucleic acids can be isolated and purified, but the extraction process takes 2-3 days and requires multiple complex steps
Solution Approach 1:
The patent combines multiple conventional extraction steps (dewaxing, cell lysis, nucleic acid release, precipitation, and purification) into a single high-pressure heating step. This merging of operations reduces the extraction process from 2-3 days to approximately 10-30 minutes while maintaining nucleic acid quality, directly resolving the time efficiency contradiction
Solution Approach 2:
The patent changes the physical parameters of the extraction system by applying high pressure (1-10 MPa) and elevated temperature (100-200°C) simultaneously. These parameter changes enable the breakdown of paraffin embedment and cell structures in a single step, eliminating the need for sequential conventional steps and dramatically reducing extraction time while preserving nucleic acid integrity
2Ease of manufacture
If multiple dewaxing steps in xylene and long-term digestion with high concentration proteases are performed for paraffin-embedded tissue, then paraffin can be removed and cells can be lysed, but the process time increases significantly
Solution Approach 1:
The patent merges the dewaxing, cell lysis, and nucleic acid release steps into a single high-pressure heating operation. This eliminates the need for sequential xylene dewaxing and protease digestion steps, reducing processing time from hours to minutes while maintaining the ability to handle paraffin-embedded tissues
Solution Approach 2:
The patent utilizes phase transitions of water (liquid to vapor) under high pressure to achieve simultaneous dewaxing and cell lysis. The high-pressure heating causes rapid vaporization of water, creating mechanical stress that breaks down paraffin embedment and cell membranes in a single step, dramatically reducing processing time
3Loss of time
If solid-phase nucleic acid extraction kits are used, then extraction time is shortened, but the process still requires multiple steps and specialized equipment, failing to achieve low-cost high-efficiency simple target processes
Solution Approach 1:
The patent employs a self-service approach where the high-pressure heating system performs multiple functions (dewaxing, lysis, release, precipitation) simultaneously without requiring specialized extraction kits or complex reagent sequences. The method uses simple water-based chemistry and universal laboratory equipment, eliminating the need for expensive solid-phase kits while maintaining speed and simplicity
Solution Approach 2:
The high-pressure heating method is universally applicable to various biological samples including paraffin-embedded tissues, fresh tissues, and cell cultures. A single protocol handles multiple sample types and multiple extraction objectives (dewaxing, lysis, purification) simultaneously, replacing the need for multiple specialized kits and equipment
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
This method significantly reduces the time and cost of nucleic acid extraction, enhances efficiency, and facilitates reliable nucleic acid detection from various samples, including paraffin-embedded tissues, by simultaneously dewaxing, lysing, and de-crosslinking in a single step, improving the availability of nucleic acids for genotyping and diagnosis.
Implementation Method 1
subjecting the biological sample to high-pressure heating
Implementation Method 2
subjecting the biological sample to high-pressure heating
Implementation Method 3
adding a dissolution medium into the biological sample
Implementation Method 4
centrifuging the biological sample
Data Source
AI summary
The invention discloses a method of extracting nucleic acid analyte solution from biological sample, comprising: placing the biological sample into a heating container; optionally, adding solvent medium into the biological sample; heating the biological sample under high pressure; optionally, centrifuging the biological sample; and obtaining the solution including the nucleic acid analyte. The invention also discloses a method of detecting the nucleic acid analyte obtained by said extraction method.