Nucleic Acid Probe Quantitation of Mutant-to-Wild-Type Ratio
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Solution Overview
Problem
Existing methods for testing genetic mutations using DNA chips can only qualitatively determine the presence or absence of genetic mutations or quantify the abundance of nucleic acid fragments, failing to provide a quantitative analysis of the abundance ratio of the genetic mutation to the wild-type gene.
Innovation Solution
The method involves nucleic acid amplification followed by detection using complementary nucleic acid probes during the exponential amplification phase, allowing for the quantitative analysis of genetic mutations by measuring signal intensities from target and non-target nucleic acids, and utilizing a calibration curve for precise quantitation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a DNA chip system with nucleic acid probes is used for genetic mutation testing, then the presence or absence of genetic mutations can be detected, but the abundance ratio of genetic mutation to wild-type gene cannot be quantitatively analyzed
Solution Approach 1:
The invention divides the detection process into two distinct phases: (1) nucleic acid amplification phase where target and non-target sequences are amplified separately, and (2) hybridization phase where probes detect the amplified products. This segmentation allows quantitative analysis by measuring signal intensities at different stages, resolving the contradiction between measurement precision and device complexity
Solution Approach 2:
The invention performs preliminary nucleic acid amplification before hybridization detection. By amplifying the target and non-target nucleic acids in advance using specific primers, the system generates sufficient material for quantitative analysis, enabling precise measurement of abundance ratios without requiring complex direct detection methods
2Measurement precision
If nucleic acid fragments are detected at the endpoint of amplification, then the detection process is simple, but the abundance ratio of genetic mutation cannot be quantitatively measured
Solution Approach 1:
The invention uses signal intensity feedback from the hybridization step to quantify abundance ratios. By measuring the intensity of signals from target-specific probes and non-target-specific probes, and comparing these intensities, the system calculates the abundance ratio of genetic mutations, achieving precise quantitation while maintaining operational simplicity
Solution Approach 2:
The invention changes the detection parameter from binary presence/absence to continuous signal intensity measurement. By detecting the intensity of hybridization signals rather than merely detecting presence, the system can quantitatively determine abundance ratios, transforming a qualitative method into a quantitative one without significantly complicating the操作流程
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 the quantitative analysis of genetic mutations, specifically the ratio of mutant to wild-type nucleic acids, through the use of nucleic acid probes and a DNA chip system, enhancing the precision of genetic mutation testing.
Implementation Method 1
a target nucleic acid containing a base to be detected in the genetic mutation and a non-target nucleic acid containing a non-detection base corresponding to the base to be detected, which are contained in a reaction liquid in an exponential amplification phase of the nucleic acid amplification reaction, using a target nucleic acid detecting nucleic acid probe having a sequence complementary to a region including a base to be detected in the target nucleic acid
Data Source
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AI summary
A genetic mutation to be tested is quantitatively analyzed using a nucleic acid probe. A nucleic acid fragment including a genetic mutation is detected using the nucleic acid probe in an exponential amplification phase of a nucleic acid amplification reaction.