Wheat DNA Detection Using Single-Copy PCR Primers
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting and quantifying endogenous wheat DNA in food samples are hindered by high homology with other cereals like barley and rye, leading to false detection and inaccurate quantification of genetically modified (GM) wheat contamination, especially since GM wheat has not yet undergone safety assessment and specific PCR primer pairs are lacking.
Innovation Solution
Development of PCR primer pairs targeting specific single-copy regions of wheat DNA, such as the WaxyD, TaSUT1D, CbpIII, GSS, and Lr1 gene regions, which are unique to wheat and do not cross-react with other crops, allowing for accurate detection and quantification of endogenous wheat DNA without false positives.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional PCR methods using general cereal primers are used, then detection of wheat DNA is possible, but false positives occur due to high homology with other cereals like barley and rye
Solution Approach 1:
The patent applies local quality by designing PCR primers that target specific unique regions within the wheat genome (WaxyD, TaSUT1D, CbpIII, GSS, and Lr1 genes) rather than using general cereal primers. This localized targeting ensures that only wheat-specific DNA sequences are amplified, eliminating cross-reactivity with other cereals while maintaining detection sensitivity.
2Measurement precision
If multi-copy DNA regions are targeted for PCR amplification, then detection sensitivity is improved, but quantification accuracy decreases due to inability to determine copy number
Solution Approach 1:
The patent changes the fundamental parameter of target selection from multi-copy regions to single-copy regions in the wheat genome. By selecting genes known to exist as single copies (WaxyD, TaSUT1D, CbpIII, GSS, and Lr1), the method establishes a direct 1:1 relationship between detected PCR products and wheat genome copies, enabling accurate quantification of wheat DNA in food samples.
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 enables precise detection and quantification of wheat in food samples, preventing false positives and ensuring accurate assessment of GM wheat contamination by utilizing primer pairs that specifically target wheat DNA sequences present in single copies, thereby enhancing the reliability of GM wheat contamination analysis.
Implementation Method 1
Methods of detecting GMOs in test samples of processed foods and their raw materials, etc. include methods of detecting modified DNA by polymerase chain reaction (PCR)
Data Source
AI summary
A circular DNA is provided comprising endogenous DNA common to both genetically modified wheat and non-genetically modified wheat along with one or more pieces of DNA each having a sequence present specifically in a strain of genetically modified wheat. Also provided is a method for determining a mix rate of genetically modified wheat in a test sample.