Assist Probe Design for High-Sensitivity Gene Detection
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Solution Overview
Problem
Current signal amplification methods using oligonucleotides for detecting genes, such as the PALSAR method, face challenges in increasing sensitivity and simultaneously detecting multiple genes effectively.
Innovation Solution
Designing a specific assist probe with nucleic acid regions X, Y, and Z, and a target region, capable of hybridizing with a target gene, along with spacer regions to enhance binding and sensitivity, allowing for the formation of a signal probe polymer that can detect multiple genes by varying the target regions of the assist probes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If an assist probe with a sequence complementary to one region in an HCP is used, then the method can detect a target gene, but the detection sensitivity is insufficient
Solution Approach 1:
The assist probe is divided into multiple functional regions: a target gene binding region (complementary to the target gene) and multiple HCP binding regions (complementary to different regions in the HCP). This segmentation allows the probe to simultaneously bind to both the target gene and multiple HCP molecules, forming a stable complex that enhances detection sensitivity while maintaining reliability
Solution Approach 2:
The assist probe is designed with multi-functionality by incorporating multiple HCP binding regions that can interact with different parts of the HCP structure. This allows a single assist probe design to work with various HCP sequences and target genes, improving both detection sensitivity through multiple binding interactions and universality across different detection applications
2Adaptability or versatility
If multiple assist probes with different target regions are prepared to detect multiple genes, then gene detection versatility increases, but the complexity of probe preparation and reaction time increase
Solution Approach 1:
The assist probe incorporates universal HCP binding regions that can interact with conserved sequences in different HCP molecules. This universal binding capability allows the same assist probe structure to detect multiple different target genes by simply changing the target gene binding region sequence, thereby achieving multi-gene detection without proportionally increasing system complexity
Solution Approach 2:
The assist probe is pre-designed with optimized structures including multiple HCP binding regions and appropriate spacer sequences before the detection reaction. This preliminary optimization of probe structure ensures that when multiple assist probes are used for different genes, they all function efficiently with consistent binding kinetics, reducing the complexity of reaction condition optimization and speeding up the overall detection process
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
Significantly increases the sensitivity of gene detection and enables the simultaneous detection of various genes by optimizing the assist probe structure and reaction processes.
Implementation Method 1
an assist probe having a plurality of the same nucleic acid regions as in the first probe and a target region capable of hybridizing with a target gene
Implementation Method 2
a signal amplification method using a pair of oligonucleotides that form a self-assembly substance (polymer) of the HCPs
Data Source
AI summary
Provided are a method of detecting target genes capable of increasing sensitivity in a Palsar method and of simultaneously detecting multiple genes, an assist probe to be used in the above method, and a method of forming a signal probe polymer by using the assist probe. The method of detecting target genes includes: forming a signal probe polymer by using a first probe having a nucleic acid region X, a nucleic acid region Y and a nucleic acid region Z in the stated order from the 5′ end, a second probe having a nucleic acid region X′, a nucleic acid region Y′ and a nucleic acid region Z′ in the stated order from the 5′ end, and an assist probe having a plurality of the same nucleic acid regions as in the first probe and a target region capable of hybridizing with a target gene. In this method, the assist probes are designed so as to have a structure including the nucleic acid regions X, Y and X, and the target region in the stated order from the 5′ end or a structure including the target region, and the nucleic acid regions Z, Y and Z in the stated order from the 5′ end.