Membrane-Coupled Nanopore Detection for Ultra-Low DNA Sequencing
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Solution Overview
Problem
Existing nucleic acid sequencing technologies are slow and expensive due to reliance on amplification techniques and require high concentrations of fluorescent chemicals, limiting their application in low-concentration samples like those from cancer cells.
Innovation Solution
Coupling the analyte to a membrane with a detector present on the same plane, allowing for ultra-low concentration detection by increasing capture frequency and reducing interference with nanopore-based sequencing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If amplification techniques are used to produce large volumes of nucleic acid, then detection sensitivity is improved, but the complexity and cost of the sequencing process increases
Solution Approach 1:
The invention extracts and eliminates the amplification step from the sequencing process by using nanopore detection that can directly detect single molecules of nucleic acid at ultra-low concentrations (0.001pM), thereby simplifying the overall sequencing workflow while maintaining high detection sensitivity
Solution Approach 2:
The invention changes the detection parameter from requiring high concentrations of fluorescent chemicals to detecting electrical current blockades at the single-molecule level, enabling direct detection without amplification and reducing process complexity
2Measurement precision
If high concentrations of fluorescent chemicals are used for signal detection, then detection accuracy is improved, but the cost and environmental impact increases
Solution Approach 1:
The invention substitutes optical detection (fluorescence) with electrical detection (current blockade measurement) at the nanopore level, eliminating the need for fluorescent chemicals while maintaining high detection accuracy through electrical signal measurement
Solution Approach 2:
The invention changes the detection modality from optical fluorescence requiring high chemical concentrations to electrical current measurement at the single-nanopore level, thereby detecting nucleic acids at ultra-low concentrations without fluorescent chemicals
3Productivity
If polynucleotide concentration is increased to maximize enzyme-analyte interactions, then sequencing efficiency is improved, but the applicability to dilute samples like cancer cell DNA is reduced
Solution Approach 1:
The invention enables the system to function effectively at ultra-low concentrations by using the nanopore itself as the detection element that can capture and detect single molecules, eliminating the need for high analyte concentrations to drive enzyme-analyte interactions
Solution Approach 2:
The invention changes the operating concentration parameter from nanomolar/ranomolar ranges required for conventional sequencing to picomolar/femtomolar ranges (0.001pM) for nanopore detection, expanding adaptability to dilute clinical samples while maintaining sequencing efficiency
4Reliability
If DNA is tethered to the membrane away from the pore, then permanent blocking is prevented, but capture efficiency may be reduced
Solution Approach 1:
The invention uses dynamically controllable tethers that can switch between tethered and untethered states, allowing the DNA to be captured efficiently when free in solution and then held in position for sequencing without permanent blocking of the pore
Solution Approach 2:
The invention employs periodic or controllable tethering where the DNA is temporarily held away from the pore during sequencing to prevent blocking, then released for capture, creating a cyclical pattern that maintains both pore availability and capture efficiency
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
Achieves ultra-sensitive detection of nucleic acids at concentrations as low as 0.001pM, enhancing sequencing efficiency and reducing sample preparation requirements for diagnostic devices.
Implementation Method 1
When a potential is applied across a nanopore, there is a drop in the current flow when an analyte, such as a nucleotide, resides transiently in the barrel for a certain period of time. Nanopore detection of the analyte gives a current blockade of known signature and duration.
Implementation Method 2
Coupling the analyte to a membrane with a detector present on the same plane, allowing for ultra-low concentration detection by increasing capture frequency
Data Source
Figure 1A~1B
Figure 2A~2D
Figure 3
AI summary
The invention relates to a new method of determining the presence, absence or characteristics of an analyte. The analyte is coupled to a membrane. The invention also relates to nucleic acid sequencing.