Microcarrier Multiplex DNA Mutation Detection Assay
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Solution Overview
Problem
Current diagnostic techniques for detecting DNA mutations associated with cancer are often expensive, time-consuming, and lack the ability to simultaneously analyze multiple gene mutations with high accuracy.
Innovation Solution
A method using microcarriers encoded with identifiers and coupled with probes specific for DNA mutations in the KRAS, BRAF, CTNNB1, and APC genes, allowing for multiplex detection of these mutations through PCR amplification and hybridization.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple singleplex assays are performed in parallel or sequentially to detect multiple DNA mutations, then each target can be analyzed individually, but the process becomes cumbersome and requires large sample volumes
Solution Approach 1:
The patent combines multiple singleplex assays into a single multiplex assay by immobilizing multiple different capture agents on the same solid phase surface, allowing simultaneous detection of multiple DNA mutations in one reaction vessel, thereby reducing assay complexity and sample volume requirements while maintaining detection accuracy
Solution Approach 2:
The solid phase surface is designed to perform multiple functions by supporting various capture agents that can bind to different target sequences, enabling a single assay platform to detect multiple different DNA mutations simultaneously rather than requiring separate assays for each target
2Adaptability or versatility
If multiple singleplex assays are performed to examine multiple genes simultaneously, then comprehensive analysis can be achieved, but large sample volumes are required
Solution Approach 1:
Multiple capture agents targeting different genes are merged onto a single solid phase surface, enabling comprehensive analysis of multiple genes in one reaction vessel with dramatically reduced sample volume requirements compared to performing separate singleplex assays
3Measurement precision
If conventional diagnostic techniques are used to detect DNA mutations, then individual gene testing can be performed, but the process is expensive and time-consuming
Solution Approach 1:
The patent merges multiple mutation detection assays into a single multiplex assay, allowing simultaneous analysis of multiple genes in one reaction, which dramatically increases diagnostic efficiency and reduces both time and cost while maintaining high mutation detection accuracy through specific capture agents and detection molecules
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 enables cost-effective, rapid, and accurate multiplex screening for DNA mutations, reducing the need for invasive sample retrieval and improving diagnostic efficiency.
Implementation Method 1
agents capable of specifically capturing the target macromolecules are attached to a solid phase surface. These immobilized molecules may be used to capture the target macromolecules from a complex sample by various means, such as hybridization (e.g., in DNA, RNA based assays)
Data Source
Figure 1A~1B
Figure 1C~1D
Figure 2A~2B
AI summary
Provided herein are methods and kits for detecting the presence of DNA mutations in the KRAS, BRAF, CTNNBl, and APC genes. The methods and kits employ microcarriers, each with a probe specific for a DNA mutation in the KRAS, BRAF, CTNNBl, or APC gene and an identifier unique to the probe sequence. Upon isolation and amplification of DNA from a sample, hybridization of amplified DNA with a probe, specific for a DNA mutation, that is coupled to a microcarrier indicates the presence of the DNA mutation in the sample. Since each microcarrier can be identified through detection of the identifier, multiplex screening assays for multiple mutations in each of the KRAS, BRAF, CTNNBl, and APC genes are provided.