Wheat Detection via Genome-Specific PCR Primers
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Solution Overview
Problem
Current methods fail to accurately and sensitively discriminate between common wheat and durum wheat in processed food products, as their protein components are similar, and existing PCR-based detection methods are not suitable for quantitative measurements due to DNA fragmentation during food processing.
Innovation Solution
Designing a primer set and nucleic acid probe specific to the starch synthase II-D gene on the wheat D genome, combined with a method targeting the common region of the A, B, and D genomes, to enable specific and quantitative detection of common wheat using PCR, allowing for discrimination between common wheat and durum wheat.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If protein-based detection methods are used to discriminate common wheat and durum wheat, then the detection can be performed using available commercial equipment, but the discrimination accuracy is insufficient due to similar protein compositions
Solution Approach 1:
The patent replaces protein-based detection methods with DNA-based PCR detection. Instead of analyzing protein components that are similar between common wheat and durum wheat, the invention uses specific DNA sequences (Wx-D1 gene for common wheat, SSII-D gene for durum wheat) that provide unique genetic fingerprints, enabling accurate discrimination at the molecular level.
Solution Approach 2:
The patent changes the detection parameter from protein composition to specific DNA sequence presence/absence. By targeting genome-specific markers (D genome for common wheat, A and B genomes for durum wheat), the method transforms the detection approach to use genetic parameters that fundamentally differ between the two wheat types, ensuring high discrimination accuracy.
2Measurement precision
If existing PCR-based detection methods targeting common wheat regions are used, then high sensitivity detection is achieved, but quantitative measurement is not suitable due to DNA fragmentation during food processing
Solution Approach 1:
The patent divides the detection approach into two separate PCR assays: one targeting common wheat-specific sequences (Wx-D1 gene) and another targeting durum wheat-specific sequences (SSII-D gene). This segmentation allows each assay to be optimized for its specific target, ensuring both high sensitivity for common wheat detection and reliability for quantitative measurement of durum wheat content in processed foods.
Solution Approach 2:
The patent uses specific genomic markers as intermediaries to indirectly detect the presence and quantity of each wheat type. Instead of directly measuring the wheat materials themselves, the method detects specific DNA sequences (Wx-D1 for common wheat, SSII-D for durum wheat) that serve as reliable proxies, enabling accurate quantitative measurement even when the food has undergone processing that may alter the physical state of the wheat components.
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 achieves highly sensitive and specific detection of common wheat, enabling accurate qualitative and quantitative differentiation from durum wheat, even in processed foods, with a detection limit of 50 pg/μL, and is suitable for use in food labeling and quality control.
Implementation Method 1
implementing a PCR procedure using a primer having the base sequence shown by SEQ ID NO:5 and a primer having the base sequence shown by SEQ ID NO:6, with a nucleic acid extracted from the sample of interest being used as a template; and detecting the presence of a PCR amplification product
Implementation Method 2
Designing a primer set and nucleic acid probe specific to the starch synthase II-D gene on the wheat D genome, combined with a method targeting the common region of the A, B, and D genomes
Data Source
AI summary
Disclosed are: a method for detecting common wheat among from wheat varieties contained in a sample of interest such as a food raw material or a processed food specifically, with high sensitivity, and in a qualitative and/or quantitative manner; a method for discriminating between common wheat and a wheat variety other than common wheat (e.g., durum wheat) contained in a food raw material or a processed food and detecting the common wheat in a qualitative and/or quantitative manner; and a primer set, a nucleic acid probe, and a detection kit, each of which can be used in the methods employing a PCR method. Specifically disclosed are: a method for detecting the occurrence of common wheat in a sample of interest, which comprises carrying out a PCR method using a nucleic acid extracted from the sample as a template and using a primer comprising the nucleotide sequence represented by SEQ ID NO:5 and a primer comprising the nucleotide sequence represented by SEQ ID NO:6 and detecting the occurrence of a PCR amplification product; and a method for detecting the occurrence of common wheat in a sample of interest, which comprises carrying out a quantitative PCR method using a nucleic acid extracted from the sample as a template and using a primer comprising the nucleotide sequence represented by SEQ ID NO:5, a primer comprising the nucleotide sequence represented by SEQ ID NO:6 and a nucleic acid probe comprising the nucleotide sequence represented by SEQ ID NO:11 and detecting the occurrence of common wheat qualitatively and/or quantitatively.