Graphene FET Biosensor for Single Nucleotide Mismatch Detection

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Solution Overview

Problem

Current methods for detecting single nucleotide polymorphisms (SNPs) in nucleic acids face challenges such as high costs, long processing times, and reliability issues, particularly with enzyme-based methods, and hybridization-based methods struggle with cross-hybridization and specificity, especially with longer probe lengths, limiting the detection of single mismatches in longer DNA strands.

Innovation Solution

The development of an electro-based nucleic acid detection system using a graphene field-effect transistor (FET) biosensor that measures changes in electric current and resistance to detect single nucleotide polymorphisms, employing DNA zippers or tweezers with toehold-mediated strand displacement, and a fluorescence-based method to discriminate between fully complementary and mismatched nucleic acid sequences.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Measurement precision

If enzyme-based methods are used for SNP detection, then detection capability is improved, but cost and processing time increase significantly

Engineering Contradiction:
ImproveSNP detection capabilityVSAvoidprocessing time
Core Design Contradiction:
Measurement precisionVSLoss of time

Solution Approach 1:

The patent replaces enzyme-based biochemical systems with a solid-state graphene FET sensor system. The graphene field-effect transistor directly transduces nucleic acid hybridization events into electrical signals, eliminating the need for enzymatic amplification steps and associated processing time while maintaining high detection precision for single nucleotide polymorphisms.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The invention changes the detection parameter from optical or biochemical readouts to electrical conductance measurements. By monitoring changes in electrical current through the graphene channel upon nucleic acid binding, the system achieves rapid detection without the time-consuming enzymatic reactions required by traditional methods.

Inventive Principle:
Principle #35Parameter changes

2Quantity of substance

If longer probe lengths are used in hybridization-based methods, then detection coverage is improved, but specificity decreases due to cross-hybridization

Engineering Contradiction:
Improveprobe lengthVSAvoiddetection specificity
Core Design Contradiction:
Quantity of substanceVSReliability

Solution Approach 1:

The patent employs local quality by using shorter, highly specific probe sequences that are optimized for individual SNP detection sites. Rather than relying on long probes that may cross-hybridize, the system uses targeted short probes that maintain high specificity while the graphene FET's high sensitivity compensates for the reduced probe length through enhanced signal transduction.

Inventive Principle:
Principle #3Local quality

Solution Approach 2:

The replacement of traditional hybridization-based detection with graphene FET electrical sensing allows the use of shorter probes. The graphene sensor's high surface area to volume ratio and sensitive electron transport properties enable reliable detection even with reduced probe lengths, eliminating the cross-hybridization problems inherent in long-probe methods.

Inventive Principle:
Principle #28Mechanics substitution (Replace mechanical system)

3Reliability

If traditional biosensors are used, then detection functionality is achieved, but sensitivity is insufficient for single mismatch detection

Engineering Contradiction:
Improvedetection functionalityVSAvoidsensitivity
Core Design Contradiction:
ReliabilityVSMeasurement precision

Solution Approach 1:

The patent utilizes graphene, a two-dimensional carbon material with exceptional electrical properties, as the sensing element. Graphene's high electron mobility, large surface area, and sensitivity to surface charge changes enable the detection of single nucleotide mismatches through minute changes in electrical conductance, providing sensitivity far exceeding traditional silicon-based or other conventional biosensor materials.

Inventive Principle:
Principle #40Composite materials

Solution Approach 2:

The invention changes the fundamental detection parameter by using electrical conductance measurements instead of optical or other traditional readouts. This parameter change, combined with graphene's inherent properties, amplifies the signal from single mismatch events, achieving the required sensitivity for discriminating single nucleotide polymorphisms in long DNA sequences.

Inventive Principle:
Principle #35Parameter changes

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 enables sensitive and specific detection of single nucleotide mismatches in long DNA sequences, enhancing sensitivity by up to -200,000-fold compared to current devices, facilitating portable and cost-effective detection of nucleic acid sequences, suitable for in-field or at-home diagnostics.

Implementation Method 1

graphene FET biosensor that measures changes in electric current and resistance

Methodology Applied
Scientific EffectField-effect transistor sensing: Electric Field

Implementation Method 2

measures changes in electric current and resistance

Methodology Applied
Scientific EffectElectrical resistance: Electrical Resistance

Implementation Method 3

employing DNA zippers or tweezers with toehold-mediated strand displacement

Methodology Applied
Scientific EffectStrand displacement:

Implementation Method 4

hybridization of a probe nucleic acid and a target nucleic acid

Methodology Applied
Scientific EffectHybridization:

Implementation Method 5

transmitted remotely using the microcontroller board and communicated in real time by wireless communication via Bluetooth standard

Methodology Applied
Scientific EffectWireless communication: Electromagnetic Induction

Data Source

PatentEP3402745B1NANO-sensors for nucleic acid detection and discrimination
Publication Date: 2023.07.26 RGT UNIV OF CALIFORNIA
  • EP3402745B1 patent drawingFigure 1
  • EP3402745B1 patent drawingFigure 2
  • EP3402745B1 patent drawingFigure 3

AI summary

Methods, systems, and nano-sensor devices are disclosed for detecting or discriminating nucleic acids with a single nucleotide resolution based on nucleic acid strand displacement. The detection can be made by time-lapse fluorescence measurements or by electro-based graphene FET, which can be combined with wireless communication to provide real-time transmission of the detected signals.