Unstable cfDNA Detection Using Charged Nanowires Without PCR
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Solution Overview
Problem
Current methods for detecting cell-free DNA (cfDNA) in liquid biopsies require amplification processes like PCR, which are cumbersome and limit on-site diagnosis, and there is a need for improved sensitivity and accuracy in detecting unstable cfDNA for cancer diagnosis.
Innovation Solution
A method and device for detecting unstable cfDNA without amplification by using positively charged nanowires and probes that bind to unstable cfDNA with a marker, allowing quick and accurate detection of gene mutations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If PCR amplification is used to detect cfDNA, then detection sensitivity is improved, but device complexity and analysis time increase
Solution Approach 1:
The patent extracts and utilizes the inherent instability of cfDNA from tumor cells as a direct detection target, removing the need for PCR amplification. By detecting structural characteristics (nicks, gaps, breaks) of unstable cfDNA through specific binding proteins and imaging, the method achieves cancer detection without complex amplification equipment
Solution Approach 2:
The patent replaces the mechanical/chemical PCR amplification system with a direct structural detection system. Instead of using polymerase enzymes and thermal cycling to amplify DNA, the method uses specific binding proteins that recognize and bind to structural defects in unstable cfDNA, followed by direct imaging detection
2Measurement precision
If PCR amplification is used to detect cfDNA, then detection sensitivity is improved, but analysis time is increased
Solution Approach 1:
The patent performs preliminary enrichment of unstable cfDNA from the complex biological sample before detection. By using specific binding proteins that selectively bind to structural defects in tumor-derived cfDNA, the method pre-concentrates the target molecules, enabling direct detection without time-consuming amplification cycles
Solution Approach 2:
The patent skips the entire PCR amplification process and goes directly from cfDNA extraction to structural detection. By rushing through the intermediate amplification step and directly detecting the inherent structural characteristics of unstable cfDNA, the method dramatically reduces analysis time while maintaining detection sensitivity
3Measurement precision
If amplification processes are used for cfDNA detection, then detection accuracy is improved, but ease of operation is reduced
Solution Approach 1:
The patent enables the detection system to self-differentiate between stable and unstable cfDNA based on their intrinsic structural properties. The binding proteins automatically recognize and bind to nicks, gaps, or breaks in the DNA backbone without requiring manual intervention or complex protocol steps, making the assay easy to perform while maintaining high accuracy
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 rapid and precise identification of unstable cfDNA from small biological samples, facilitating early cancer diagnosis and prognosis by eliminating the need for amplification processes and reducing analysis time.
Implementation Method 1
mixing a sample containing cfDNA with a positively charged nanowire
Implementation Method 2
probes that bind to unstable cfDNA with a marker, allowing quick and accurate detection of gene mutations
Data Source
AI summary
In an embodiment, the present invention relates to a technique in which small-sized cfDNA is detected, with ultrahigh sensitivity, from a liquid sample such as urine, cerebrospinal fluid, plasma, blood, pleural fluid, or body fluid, is concentrated and isolated, and then is analyzed for gene mutations without PCR. In particular, in a case where a positively charged nanostructure is used, capture and detection rates for cfDNA can be increased. A detection method according to an embodiment of the present invention does not require a PCR amplification reaction, which greatly shortens the time taken to obtain a result. In addition, since direct on-site analysis is possible without the need for specific equipment, it is expected that the present invention can be used as a point-of-care testing (POCT) capable of simultaneously searching multiple genes in a short period of time.