Electrode Binary Probe for Nucleic Acid Quantification and Mutation Detection
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Solution Overview
Problem
Existing electrochemical methods are unable to simultaneously quantify the concentration of specific nucleic acid targets and detect mutations in nucleic acid sequences on the same platform.
Innovation Solution
A binary probe composed of long and short nucleic acid chains tethered to an electrode, where the short chain has a covalently bonded redox active compound, is used to measure the binding of specific nucleic acid targets, allowing for toe-hold displacement and electrochemical signal change to determine concentration and mutation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If existing electrochemical methods are used, then measurement of nucleic acid targets is possible, but simultaneous quantification of concentration and detection of mutations cannot be achieved
Solution Approach 1:
The binary probe system performs multiple functions simultaneously: it quantifies nucleic acid concentration through electrochemical signal changes and detects mutations through kinetic analysis of binding rates. The same probe structure (short chain tethered to electrode with long chain capable of toe-hold displacement) enables both concentration measurement and mutation detection without requiring separate systems
Solution Approach 2:
The probe is divided into functional segments: a short chain tethered to the electrode providing electrochemical signal, and a long chain with toe-hold displacement capability for specific target binding. This segmentation allows the short chain to mediate electrochemical signals while the long chain performs sequence-specific recognition and mutation detection
2Reliability
If multiple nucleic acid sequences are analyzed for complex diseases, then reliable disease conclusion can be obtained, but the complexity of analysis increases
Solution Approach 1:
The binary probe system can be configured to detect multiple different target sequences by changing the probe sequence. The same electrochemical platform enables analysis of multiple nucleic acid sequences for complex diseases, providing reliable conclusions through integrated concentration and mutation detection for each target
Solution Approach 2:
The method uses sequence-specific probes that can be designed to match different target sequences. By creating probe copies with different sequences, the system can simultaneously or sequentially analyze multiple nucleic acid targets for complex disease diagnosis without requiring fundamentally different analytical approaches
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
The method enables quantitative analysis of nucleic acid concentrations and detection of mutations by measuring electrochemical signals, providing a unified platform for both measurements.
Implementation Method 1
The other end of the short chain, subsequently called the free end, has a covalently bonded redox active compound (RAC). Subsequently, the RAC on the released short chain can undergo redox causing the electrochemical a signal to increase.
Implementation Method 2
The long chain is bound to the short chain by hydrogen bonding over the complimentary sequence to form the well-known double helix structure.
Data Source
AI summary
A binary probe composed of long and short nucleic acid (NA) or peptide nucleic acid (PNA) chains immobilized on an electrode to electrochemically detect specific sequences of single stranded nucleic acids extracted from biospecimens by specific binding to a single spot or and an array of spots of the probe molecules. The short chain is tethered to the electrode at one end and the other end has a covalently bound redox active compound (RAC). Part of the long chain is complimentary to the short chain to form a hydrogen bonded duplex. On specific binding of the long chain to the specific target of interest the electrochemical signal form the RAC changes due to release of the long chain. The potential applicability of the invention is in genomics to quantify certain target sequences and detect mutations for medical diagnosis, track efficacy of therapy, fundament research in life sciences, and drug development.