Somatic Mutation Detection via Peripheral Blood Mononuclear Cell DNA
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Solution Overview
Problem
Current methods for detecting somatic mutations in tumor marker genes from tumor tissues are invasive and have low accuracy, especially when using blood samples, as the small amount of cfDNA in blood serum or plasma makes it difficult to detect mutations reliably.
Innovation Solution
The method involves recovering the peripheral blood mononuclear cell layer from a blood sample and using DNA from this layer to detect somatic mutations in tumor marker genes using techniques like real-time PCR, digital PCR, or next-generation sequencing, which increases detection sensitivity and accuracy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If cfDNA in blood plasma or serum is used for detecting somatic mutations, then the detection can be performed non-invasively, but the detection accuracy is insufficient due to the extremely small amount of cfDNA (approximately 10³ copies) and low allele proportion (approximately 0.1%)
Solution Approach 1:
The invention extracts and isolates cfDNA from blood plasma or serum through centrifugation and purification steps, concentrating the target DNA from a complex biological matrix. This extraction process separates cfDNA from proteins, cells, and other interfering substances, enabling subsequent sensitive detection of somatic mutations despite the low abundance of target molecules.
Solution Approach 2:
The invention employs PCR amplification to exponentially increase the concentration of target DNA sequences containing somatic mutations. By using specific primers that bind to mutant alleles, the method amplifies only the relevant sequences, transforming the undetectably low initial concentration into a measurable amount while maintaining the non-invasive nature of the original sample collection.
2Measurement precision
If tumor tissue samples are collected by biopsy to examine genotypes of cancer genes, then the detection accuracy is high, but the procedure causes extreme invasion and discomfort to patients
Solution Approach 1:
The invention uses cfDNA in blood plasma or serum as an intermediary substance that carries genetic information from tumor tissues without requiring direct access to the tumor itself. This liquid biopsy approach allows tumor genotype analysis through a peripheral blood sample, eliminating the need for invasive tissue biopsy while preserving detection accuracy through targeted molecular analysis of the circulating DNA.
3Loss of time
If chronological biopsy is performed to monitor somatic mutations over time, then real-time tumor status can be understood, but repeated biopsies are difficult to perform due to the invasive nature
Solution Approach 1:
The invention enables continuous monitoring of somatic mutations through repeated blood draws, which can be performed frequently without significant patient burden. The cfDNA in plasma or serum provides a continuous source of tumor genetic information that can be analyzed over time, allowing chronological tracking of mutation dynamics, treatment response, and tumor evolution without the limitations of repeated tissue biopsies.
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 allows for the high-sensitive detection of somatic mutations in tumor marker genes, enabling accurate evaluation of tumor status and drug susceptibility with reduced invasiveness, even when mutations are not detectable in blood plasma or serum.
Implementation Method 1
recovering a peripheral blood mononuclear cell layer from a blood sample
Data Source
AI summary
A method of detecting somatic mutations of tumor marker genes includes recovering a peripheral blood mononuclear cell layer from a blood sample collected from a subject and examining whether or not a nucleic acid derived from tumor marker genes having somatic mutations is detected from DNA included in the peripheral blood mononuclear cell layer.