Nucleic Acid Quality Determination via Internal Quality Control Index
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Solution Overview
Problem
Current methods for analyzing nucleic acid quality in biological samples, particularly in FFPE tissues, often result in inaccurate mutation analysis due to DNA fragmentation and cross-linking, leading to unreliable results in genetic diagnostics and companion diagnostics.
Innovation Solution
A method involving the use of primers capable of amplifying an internal quality control region through PCR, calculating the internal quality control index, and standardizing mutation frequency by dividing measured mutation frequency by this index, ensuring the integrity and amplifiability of nucleic acid templates.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional PCR methods are used to analyze nucleic acids in biological samples, then the analysis can be performed, but the results are inaccurate due to DNA fragmentation and cross-linking
Solution Approach 1:
The method performs preliminary quality assessment of nucleic acid templates before mutation analysis by calculating the internal quality control index. This preliminary action identifies samples with insufficient DNA integrity or amplifiability, allowing them to be excluded or pre-treated before the main mutation analysis, thereby preventing inaccurate results
Solution Approach 2:
The invention introduces an intermediary quality control index (iQC index) that mediates between the raw nucleic acid sample and the mutation analysis process. This index serves as a bridge to objectively evaluate template quality and standardize mutation frequency data across different sample qualities
2Ease of manufacture
If mutation analysis is performed without quality standardization, then the analysis process is simple, but the results vary between laboratories
Solution Approach 1:
The method changes the parameter of mutation frequency reporting by introducing a standardized mutation frequency that is adjusted based on the internal quality control index. This parameter transformation allows results to be corrected for differences in DNA quality and amplifiability, enabling consistent comparison across laboratories while maintaining relatively simple processing
3Productivity
If nucleic acid quality is not evaluated, then the analysis time is short, but the accuracy of genetic diagnostics is compromised
Solution Approach 1:
The quality evaluation using the internal quality control index is performed as a preliminary step before mutation analysis. This preliminary assessment quickly identifies unsuitable samples, allowing rapid exclusion of poor-quality samples without requiring complex additional procedures, thus maintaining productivity while improving accuracy
Solution Approach 2:
The invention replaces subjective quality assessment with an objective computational system that calculates the internal quality control index based on PCR amplification data. This substitution of mechanical/computational evaluation for subjective judgment improves accuracy while requiring minimal additional time
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 approach objectively evaluates nucleic acid quality and provides reliable mutation frequency data, enhancing the accuracy of genetic analysis and companion diagnostics by accounting for DNA integrity and amplifiability.
Implementation Method 1
performing PCR on the extracted nucleic acids with i) primers; or ii) a set of primers and a probe which are capable of amplifying the internal quality control region
Data Source
AI summary
The present invention relates to a method for determining the DNA quality of a biological sample and, more specifically, to a method for determining the DNA quality of a biological sample by performing a quantitative polymerase chain reaction (PCR) using primers capable of amplifying a target gene, a method for preparing the primers used in the method, and a method for standardizing the amount of detected target gene mutation by using the determined DNA quality. The method of the present invention enables objective evaluation of the DNA quality of a biological sample used in gene analysis and the presentation of objective results on the expression ratio of a gene mutation, thereby providing reliable information in the fields of clinical research and companion diagnosis.