EDTA Pretreatment Composition for Rapid Environmental Nucleic Acid Detection
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for nucleic acid detection in environmental samples, such as water, are cumbersome, time-consuming, and costly, and are susceptible to nuclease interference, necessitating a more efficient and accurate pretreatment process.
Innovation Solution
A method involving the use of ethylenediaminetetraacetic acid (EDTA) for pretreatment, combined with heat treatment at 90°C or higher, to stabilize nucleic acids and inhibit nuclease activity, followed by rapid nucleic acid detection using RT-PCR or PCR.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If commercial nucleic acid extraction kits are used, then nucleic acid extraction can be performed, but the process takes approximately 1 hour and is complicated
Solution Approach 1:
The invention extracts and removes nucleic acids from the sample through a simplified heat treatment process, separating the nucleic acid extraction function from complex commercial kits. By using selective heat treatment at specific temperatures, the method isolates nucleic acids from contaminants without requiring multi-step commercial extraction procedures.
Solution Approach 2:
The invention changes the temperature parameter to achieve nucleic acid extraction. By controlling heat treatment at specific temperatures (e.g., 95-100°C for 5-10 minutes), the method transforms the physical state of nucleic acids to enable their extraction, replacing the complex chemical reagents and multiple steps of commercial kits with a single parameter-based process.
2Measurement precision
If nucleic acid extraction is performed to detect viral load, then accurate detection can be achieved, but the process is time-consuming and costly
Solution Approach 1:
The invention performs preliminary heat treatment to extract and stabilize nucleic acids before the actual detection process. This preliminary action prepares the sample in advance, ensuring nucleic acids are readily available for detection while preventing degradation by nucleases during the extraction process.
Solution Approach 2:
The invention introduces EDTA as an intermediary substance that chelates metal ions and inhibits nuclease activity. This intermediary protects nucleic acids from degradation during the extraction and detection process, maintaining measurement precision without requiring time-consuming commercial extraction kits.
3Stability of the object's composition
If samples are treated to inhibit nuclease activity, then nucleic acid stability is improved, but additional treatment steps are required
Solution Approach 1:
The invention merges the nucleic acid extraction function with the nuclease inhibition function into a single integrated heat treatment process. By combining these two functions that were previously separate steps, the method achieves both nucleic acid stabilization and extraction without requiring additional treatment steps.
Solution Approach 2:
The invention uses EDTA as an intermediary that simultaneously achieves multiple objectives: it chelates metal ions required for nuclease activity, inhibits nuclease degradation, and stabilizes nucleic acids during the heat treatment process. This single intermediary substance enables multiple functions without adding 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
Enables rapid, sensitive, and accurate detection of nucleic acids in environmental samples, reducing processing time to under 4 hours and minimizing nuclease interference.
Implementation Method 1
mixing a sample with a composition for pretreatment comprising ethylenediaminetetraacetic acid (EDTA)
Implementation Method 2
heating the sample mixed in step i) using a heating means maintained at 90° C. or higher
Data Source
AI summary
The present disclosure concerns detection of a target nucleic acid comprised in a sample, in particular, a water sample in the environment, such as treated sewage water or rivers, in a more convenient manner with higher accuracy. A target sample of nucleic acid detection is mixed with a composition for pretreatment comprising EDTA and the resultant is then heated using a heating means maintained at 90° C. or higher.