Electrode Array Nucleic Acid Detection via Redox Probes
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Solution Overview
Problem
Current nucleic acid detection methods, such as real-time PCR and microarray, face challenges in achieving high sensitivity and simplicity for detecting multiple target genes simultaneously.
Innovation Solution
A nucleic acid detection method involving a reaction mixture with magnesium ions (4 mM to 30 mM) and a redox probe (with an oxidation reduction potential of −0.5 V to 0.5 V) on an electrode array, where the amplification product generates an electric signal that increases with the amount of target nucleic acid, allowing for precise detection and quantification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If real-time PCR method is used for nucleic acid detection, then sensitivity is improved, but device complexity and operational complexity increase
Solution Approach 1:
The patent replaces the optical detection system of conventional real-time PCR with an electrochemical detection system. An electrode is directly placed in the reaction mixture to detect electric signals from redox probes, eliminating the need for complex optical components such as light sources, detectors, and optical pathways while maintaining high detection sensitivity.
Solution Approach 2:
The electrode serves multiple functions: it acts as both the amplification reaction container and the detection device. The reaction mixture containing the target nucleic acid, primers, polymerase, and redox probe is placed directly on the electrode, which detects the electric signals generated during amplification, thereby combining reaction and detection functions in a single component.
2Adaptability or versatility
If microarray method is used for simultaneous detection of multiple target genes, then detection capability is improved, but ease of operation deteriorates
Solution Approach 1:
The patent uses multiple electrodes arranged in an array format, where each electrode can detect different target genes simultaneously. This segmentation allows parallel detection of multiple targets while maintaining simple operation, as each electrode functions independently with its own reaction mixture containing specific primers and redox probes.
3Measurement precision
If conventional detection methods are used, then detection capability is achieved, but productivity decreases
Solution Approach 1:
The patent enables continuous monitoring of the amplification reaction by chronologically detecting electric signals from the electrode. This continuous detection allows real-time quantification of amplification products, improving throughput by eliminating the need for post-reaction processing and analysis steps required by conventional methods.
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 method enables simple, high-sensitivity detection and quantification of target nucleic acids, with the ability to detect multiple types of nucleic acids efficiently, improving precision and throughput compared to conventional techniques.
Implementation Method 1
a redox probe which has an oxidation reduction potential of −0.5 V to 0.5 V and generates a detectable electric signal of which amplitude increases with an increase in an amount of the amplification product
Data Source
AI summary
According to one embodiment, a method for detecting target nucleic acid includes the following steps. (A) A reaction field is formed by placing a reaction mixture on an electrode, and the reaction mixture contains the sample, a primer set, an amplification enzyme, 4 mM to 30 mM of magnesium ion, and a redox probe. The redox probe has an oxidation reduction potential, which generates an electric signal of which amplitude increases. (B) The reaction field is maintained under an amplification reaction condition. (C) The electric signal is detected with the electrode. (D) Existence or quantity of the target nucleic acid is determined.